OBJECTIVETo investigate the relationship between phenotype transformation and biomechanical properties of detrusor smooth muscle cell (DSMC) subjected to the cyclic mechanical stretch.

METHODSCultured rat DSMCS were grown on collagen-coated silicone membranes and subjected to continuous cycles of stretch-relaxation. All experiments were made on cells between passage 2 and 4. Each cycle consists of 5-second stretch and 5-second relaxation. The computer controlled vacuum induced 10% (I), 20% (II) and 30% (III) maximum elongation of the plate membrane at different designed pressures. We assessed DNA synthesis rate using tritiated thymidine incorporation assay. Using immunofluorescent assay and flow cytometer, we analysed the expression of SM-alpha-actin and proliferation of DSMC. The image analysis and micropipette aspiration systems were employed to investigate the single cell contraction and viscoelasticity. The elastic modulus K(1), K(2) and viscoelastic coefficient micro were determined using the three-element standard linear solid model, thus demonstrating the passive deformation ability of detrusor cells.

RESULTSAs the basic structural changes to mechanical stretch, DSMCs underwent phenotypic modulation from their normal contractile phenotype to a "synthetic" phenotype: the DSMCs became more proliferative and the actin less organized along the cell's long axis. The cell proliferation index (CPI) of control and stretched group (10%, 20%, 30% elongation) were 0.24, 0.43, 0.58 and 0.65 respectively. After mechanical stretch, the well-spread filaments changed their orientation. Contraction and viscoelasticity of single DSMC subjected to stretch both decreased significantly compared to control. The Vmax and. DeltaLmax of group III (30% elongation) saw significant decreases compared with unstretched control (P < 0.01). K(1) and K(2) decreased with the increasing of mechanical overload, however, there was no statistic difference between groups II and group III.

CONCLUSIONSStructure determines function. Conversely, dysfunction implies the structural transformation. Functional abnormalities of BOO have the structural basis: phenotype transformation of detrusor cells. Cyclic stretch and relaxation applied to DSMCs in vitro can be used to model the increases in urodynamic load experienced by the bladder detrusor muscle under the conditions of bladder outlet obstruction. Phenotypic transformation is the structural basis of functional changes of DSMC subjected to periodic overload mechanical stretch.

Animals ; Biomechanical Phenomena ; DNA ; biosynthesis ; Muscle, Smooth ; physiology ; Phenotype ; Rats ; Rats, Wistar ; Stress, Mechanical ; Urinary Bladder ; physiology ; Urinary Bladder Neck Obstruction ; physiopathology

Mycoplasma hyopneumoniae (M .hyopneumoniae) and Mycoplasma hyorhinis (M .hyorhinis) infections are common in China .To investigate the prevalence of M .hyorhinis and M .hyopneumoniae in Jiangsu Province of China ,a mo-lecular epidemiological survey was conducted from 399 nasal swab samples of unvaccinated pigs using nested Polymerase Chain Reaction (nested PCR) .Nasal swab samples were collected from Jiangquhai porcine lean (JQHPL) strain pigs and other West-ern breeds .Clinical samples were taken from each pig and divided into different groups based on ages of pigs (7 ,14 ,21 ,28 , and 35 days) .Results indicated that the prevalence of M .hyorhinis was 70 .9% from different herds in Jiangsu Province in China ,while the prevalence of M .hyopneumoniae was 13 .5% .M .hyorhinis infection was more common in pigs for less than 5 weeks of age compared to M .hyopneumoniae infection .Co-infection was also observed in 30 samples (7 .5% ) in which both M .hyorhinis and M .hyopneumoniae were detected .The M .hyorhinis infection increased as the animals grew from 7 to 35 days .The M .hyopneumoniae infection did not change significantly as the pigs grew older .Significant difference of M .hyorhi-nis infection was observed between other Western breeds and JQHPL pigs (P<0 .001) .JQHPL pigs appear to be more sensi-tive to the M .hyorhinis infection as compared to the other Western breeds .However ,there is no obvious relationship between the breed type and M .hyopneumoniae infection (P>0 .05) .

OBJECTIVESTo observe the detrusor ultrastructure in BPH patients and to investigate the relationship between detrusor instability and ultrastructure.

METHODSThe patients were divided into groups of detrusor instability(DI) and detrusor stability(DS) according to urodynamics examination. The structure of the detrusor were observed by light microscopy and transmission electron microscopy(TEM).

RESULTSThe intercellular intermediate junctions and cytoplasmic process junctions in DS were 11.34 +/- 3.23 and 4.26 +/- 1.78 respectively. The intercellular intermediate junctions decreased obviously (3.12 +/- 1.47, P < 0.01) instead of a large amount of cytoplasmic process junctions (26.37 +/- 7.14, P < 0.01) in DI.

CONCLUSIONSThere is a close relevance between intercellular junctions and DI. The observation of the ultrastructure of the detrusor is helpful for the diagnosis of BPH with DI and for the clinical treatment.

Aged ; Aged, 80 and over ; Humans ; Intercellular Junctions ; ultrastructure ; Male ; Microscopy, Electron ; Middle Aged ; Muscle, Smooth ; ultrastructure ; Prostatic Hyperplasia ; pathology ; Urinary Bladder ; ultrastructure

OBJECTIVETo investigate the outcomes of perineal urethrostomy plus secondary urethroplasty for ultralong urethral stricture and assess its influence on the patient's quality of life.

METHODSWe retrospectively analyzed 54 cases of ultralong urethral stricture treated by perineal urethrostomy from 2000 to 2010. The mean age of the patients was 40 years, and the average length of stricture was 6.5 cm. We evaluated the patients'quality of life by questionnaire investigation and the clinical outcomes based on IPSS, Qmax, the necessity of urethral dilation and satisfaction of the patients.

RESULTSThe mean Qmax of the 54 patients was (14.0 +/- 4.7) ml/min. Of the 34 cases that underwent secondary urethroplasty, 22 (64.7%) achieved a mean Qmax of (12.0 +/- 3.5) ml/min, 8 (23.5%) needed regular urethral dilatation and 4 (11.8%) received internal urethrotomy because of restenosis. IPSS scores were 5.4 +/- 2.1 and 8.5 +/- 5.8 after perineal urethrostomy and secondary urethroplasty, respectively. Fifty of the total number of patients (92.6%) were satisfied with the results of perineal urethrostomy, and 22 of the 34 (64.7%) with the results of secondary urethroplasty.

CONCLUSIONPerineal urethrostomy plus secondary urethroplasty is safe and effective for ultralong urethral stricture, and affects very little the patient's quality of life.

Adolescent ; Adult ; Aged ; Child ; Humans ; Male ; Middle Aged ; Ostomy ; Perineum ; surgery ; Quality of Life ; Retrospective Studies ; Treatment Outcome ; Urethral Stricture ; surgery ; Urologic Surgical Procedures ; methods ; Young Adult

OBJECTIVETo elucidate the details of operative technique of anastomotic posterior urethroplasty for traumatic posterior urethral strictures in attempt to offer a successful result.

METHODSWe reviewed the clinical data of 106 patients who had undergone anastomotic repair for posterior urethral strictures following traumatic pelvic fracture between 1979 and 2004. Patients'age ranged from 8 to 53 years (mean 27 years). Surgical repair was performed via perinea in 72 patients, modified transperineal repair in 5 and perineoabdominal repair in 29. Follow-up ranged from 1 to 23 years (mean 8 years).

RESULTSAmong the 77 patients treated by perineal approaches, 69 (95.8%) were successfully repaired and 27 out of the 29 patients (93.1%) who were repaired by perineoabdominal protocols were successful. The successful results have sustained as long as 23 years in some cases. Urinary incontinence did not happen in any patients while impotence occurred as a result of the anastomotic surgery.

CONCLUSIONSThree important skills or principles will ensure a successful outcome, namely complete excision of scar tissues, a completely normal mucosa ready for anastomosis at both ends of the urethra, and a tension-free anastomosis. When the urethral stricture is below 2.5 cm long, restoration of urethral continuity can be accomplished by a perineal procedure. If the stricture is over 2.5 cm long, a modified perineal or transpubic perineoabdominal procedure should be used. In the presence of a competent bladder neck, anastomotic surgery does not result in urinary incontinence. Impotence is usually related to the original trauma and rarely (5.7%) to urethroplasty.

Adolescent ; Adult ; Anastomosis, Surgical ; Child ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Urethra ; surgery ; Urethral Stricture ; etiology ; surgery ; Urologic Surgical Procedures

OBJECTIVETo study the new characteristics on diagnosis and treatment of renal tuberculosis (RT).

METHODSEighty-seven patients with renal tuberculosis were retrospectively reviewed; their diagnosis was established by standard microbiological and histological techniques.

RESULTSAtypical RT was diagnosed by various examinations, including urinary analysis, polymerase chain reaction of tuberculosis (PCR-TB), ultrasonography, intravenous urography (i.v.U), and computerized tomography (CT). Treatment consisted of antituberculous chemotherapy in all patients, in combination with nephrectomy (62.5%) or enterocystoplasty (4.6%).

CONCLUSIONSThe differential diagnosis of RT should be emphasized, especially for atypical RT, provided a much more specific diagnosis in clinical suspicion of RT. i.v.U can not be regarded as a specific examination for RT. Computerised tomography (CT) can be used for early diagnosis of RT. Surgery for RT is still ablative.

Adolescent ; Adult ; Aged ; Diagnosis, Differential ; Female ; Humans ; Male ; Middle Aged ; Tuberculosis, Renal ; diagnosis ; therapy

OBJECTIVETo understand the immunological status of Japanese encephalitis (JE) antibodies amongst migrant workers and to provide epidemiological basis for public health strategies on JE prevention and control in Shenzhen.

METHODSA multi-stage random sampling method was used, and 1003 migrant workers aged 18 to 60 from 44 factories were investigated and their serum specimens were collected. The enzyme-linked immunosorbent assay (ELISA) was used to detect JE antibodies qualitatively.

RESULTSThe gross IgG seroprevalence rate for JE was 20.2% (203/1003). Sex-specified seroprevalence was 21.2% (103/485) for male and 19.3% (100/518) for female, respectively (χ(2) = 579, P > 0.05). Age-specific seropositive rates were 22.6% (12/53) for those below 20 years old, 18.7% (120/642) for those between 20-years old, 26.0% (58/223) for those between 30-years old and 15.3% (13/85) for those on or above 40 years old (χ(2) = 7.96, P > 0.05). Proportions for self-reported positive immunization, non-immunization and unclear immunization history were 22.1% (30/136), 22.1% (51/231) and 19.2% (122/636), respectively (χ(2) = 501, P > 0.05). Seroprevalence by region of origins showed that workers from Guangdong province was the highest (30.5%, 50/164), followed by workers from Guangxi (29.7%, 22/74) whilst workers from Shan(3)xi (5.4%, 2/37) had the lowest rate. Seroprevalence rate for managers (29.0%, 31/107) was higher than that of technicians (7.1%, 1/14) (χ(2) = 21.78, P < 0.05). Serological positive rate of workers with university or above educational background was the highest (32.7%, 16/49), followed by that for individuals with college degree (10.3%, 10/97) (χ(2) = 13.02, P < 0.05).

CONCLUSIONNo associations are detected between JE seroprevalence and age, or sex, or self-reported immunization histories amongst migrant labor workers in Shenzhen. However, correlations between JE serological positive rate and region of origins, occupation and educational attainment are found to be significant. The gross seroprevalence of JE antibodies suggests that the level of JE antibodies amongst Shenzhen migrant workers is low and the population immunity barrier has yet to be established. It is necessary to strengthen prevention and control strategies of JE among labor workers of Shenzhen.

Adolescent ; Adult ; Antibodies, Viral ; blood ; China ; epidemiology ; Encephalitis, Japanese ; epidemiology ; prevention & control ; Female ; Humans ; Japanese Encephalitis Vaccines ; administration & dosage ; Male ; Middle Aged ; Transients and Migrants ; statistics & numerical data ; Young Adult

OBJECTIVETo study the prevalence of hepatitis C virus (HCV) infection and characteristics on molecular biology related to HCV among patients who were enrolled in a Methadone maintenance clinic in Wuhan.

METHODSSerum samples from 332 injection drug users (IDUs) were obtained and anti-HCV IgG was detected by enzyme linked immunosorbrent assay(ELISA), together with 86 anti-HCV positive specimens genotyped. A reverse transcriptase-polymerase chain reaction (RT-nPCR) assay using conserved primers deduced from the core-envelopel (C-E1) region of the HCV genome was employed to amplify a 474 bp fragment. Phylogenetic analysis of the C-E1 sequences was conducted by direct sequencing of the RT-nPCR products and alignment with determined by nucleotide sequencing followed by composition of a phylogenetic tree.

RESULTSThere were 313 cases (94.3%) appeared positive anti-HCV IgG in the 332 patients from a Methadone maintenance clinic in Wuhan. It was demonstrated that there were four different subtypes of HCV in that clinic in Wuhan, including 6a--71 cases (82.5%), 3b--7 cases (8.2%), 1a--5 cases (5.8%) and 1b--3 cases (3.5%).

CONCLUSIONInfection of 6a genotype HCV was predominant in patients from the Methadone maintenance clinic in Wuhan, followed by HCV 3b, 1a and 1b.

Adult ; Antibodies, Viral ; analysis ; China ; Enzyme-Linked Immunosorbent Assay ; Female ; Genotype ; Hepacivirus ; classification ; genetics ; Humans ; Male ; Methadone ; therapeutic use ; Middle Aged ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; Substance Abuse Treatment Centers ; Substance-Related Disorders ; drug therapy ; rehabilitation ; Young Adult

OBJECTIVETo observe the anti-virus effects of andrographolide (AD) on the retinoic acid-inducible gene-I (RIG-I)-like receptors (RLRs) signaling pathway when immunological cells were infected with H1N1.

METHODSLeukomonocyte was obtained from umbilical cord blood by Ficoll density gradient centrifugation, and immunological cells were harvested after cytokines stimulation. Virus infected cell model was established by H1N1 co-cultured with normal human bronchial epithelial cell line (16HBE). The optimal concentration of AD was defined by methyl-thiazolyl-tetrazolium (MTT) assay. After the virus infected cell model was established, AD was added into the medium as a treatment intervention. After 24-h co-culture, cell supernatant was collected for interferon gamma (IFN-γ) and interleukin-4 (IL-4) enzyme-linked immunosorbent assay (ELISA) detection while immunological cells for real-time polymerase chain reaction (RT-PCR).

RESULTSThe optimal concentration of AD for anti-virus effect was 250 μg/mL. IL-4 and IFN-γ in the supernatant and mRNA levels in RLRs pathway increased when cells was infected by virus, RIG-I, IFN-β promoter stimulator-1 (IPS-1), interferon regulatory factor (IRF)-7, IRF-3 and nuclear transcription factor κB (NF-κB) mRNA levels increased significantly (P<0.05). When AD was added into co-culture medium, the levels of IL-4 and IFN-γ were lower than those in the non-interference groups and the mRNA expression levels decreased, RIG-I, IPS-1, IRF-7, IRF-3 and NF-κB decreased significantly in each group with significant statistic differences (P<0.05).

CONCLUSIONSThe RLRs mediated viral recognition provided a potential molecular target for acute viral infections and andrographolide could ameliorate H1N1 virus-induced cell mortality. And the antiviral effects might be related to its inhibition of viral-induced activation of the RLRs signaling pathway.

Adaptor Proteins, Signal Transducing ; genetics ; metabolism ; Antiviral Agents ; pharmacology ; Cells, Cultured ; Coculture Techniques ; DEAD Box Protein 58 ; DEAD-box RNA Helicases ; genetics ; metabolism ; Dendritic Cells ; drug effects ; immunology ; virology ; Diterpenes ; pharmacology ; Fetal Blood ; cytology ; Humans ; Influenza A Virus, H1N1 Subtype ; drug effects ; immunology ; Influenza, Human ; drug therapy ; immunology ; virology ; Interferon-beta ; genetics ; metabolism ; Interferon-gamma ; metabolism ; Interleukin-4 ; metabolism ; Leukocytes, Mononuclear ; drug effects ; immunology ; virology ; Macrophages ; drug effects ; virology ; NF-kappa B ; genetics ; metabolism ; Promoter Regions, Genetic ; drug effects ; immunology ; RNA, Messenger ; metabolism ; Signal Transduction ; drug effects ; genetics ; immunology

OBJECTIVETo observe the expressions of the substance P (SP) mRNA and neurokinin-1 receptor (NK-1R) in the posterior horn of the L5 - S2 spinal cord in the rat model of chronic prostatitis pain, and to investigate the changes in the activation of astrocytes and influence of SP on this activation in rat spinal cord astrocytes cultured in vitro.

METHODSThe rat model of chronic prostatitis pain was established by injection of complete Freund's adjuvant (CFA) and assessed by the tail flick threshold test, the control rats injected with sodium chloride and all observed at 0, 14 and 28 days. Changes in the expressions of SP mRNA, NK-1R, glial fibrillary acidic protein (GFAP), tumor necrosis factor-alpha (TNF-alpha) and inducible nitric oxide synthase (iNOS) in the posterior horn of the L5 - S2 spinal cord were detected by RT-PCR and Western blot. Rat spinal cord astrocytes were cultured in vitro and divided into a control group, cultured with ITS cell culture fluid, and two experiment groups, with Group 1 stimulated with SP at the concentration of 10(-9) - 10(-6) mol/L for 12 hours followed by determination of the expressions of TNF-alpha, IL-1beta, NO and NOS by ELISA and nitrate reductase and colorimetric methods, and Group 2 at 10(-7) mol/L for 0, 24, 48 and 72 hours followed by detection of the GFAP expression by Western blot.

RESULTSThe expressions of SP mRNA, NK-1 R, GFAP, TNF-alpha and iNOS in the posterior horn of the L5 - S2 spinal cord were obviously higher in the rat prostatitis pain models than in the controls, successively higher at 28 than at 14 and 0 d (P < 0.01), and so was the expression of GFAP at 28 than at 14 d in the experiment groups (P < 0.05). SP induced a gradual increase at 10(-7) mol/L in the expression of GFAP in the spinal cord astrocytes at 0 -72 h, significantly different from that of the control group (P < 0.01), and it promoted the excretion of TNF-alpha and IL-1beta and the activity of NO and NOS at 10(-9) - 10(-6) mol/L at 12 h in a concentration-dependent manner, with marked differences between the experiment and control groups (P < 0.01, P < 0.05). But a decreased excretion of IL-1 beta was observed in the 10(-6) mol/L group, though with no significant difference from the control (P > 0.05).

CONCLUSIONChronic prostatitis pain could upregulate the expressions of the excitatory transmitter SP and receptor in the L5 - S2 spinal cord, and result in the activation of astrocytes and increased excretion of proinflammatory cytokines, which may be associated with the persistence and generalization of prostatitis pain.

Animals ; Astrocytes ; metabolism ; Chronic Disease ; Male ; Nitric Oxide Synthase Type II ; metabolism ; Pain ; metabolism ; Prostatitis ; metabolism ; Rats ; Receptors, Neurokinin-1 ; metabolism ; Spinal Cord ; cytology ; metabolism ; pathology ; Substance P ; metabolism