Objective To investigate the antiproliferative effect of rosiglitazone, a thiazolidinedione (TZD) on autosomal dominant polycystic kidney disease (ADPKD) cystic lining epithelial cells and to explore the underlying molecular mechanism. Methods ADPKD cysticlining immortalized epithelial (WT9-12) cells were stimulated by rosiglitazone with different concentrations. After treatment, MTT method was performed to detect the level of proliferation; flow cytometry was used to determine the cell cycle distribution and the apoptosis rate. Western blotting was used to detect the protein expressions of mTOR, p70S6K, 4E-Bp1, PPARγ PPARγ siRNA was transfected into WT9-12 cells to knock down the expression of PPARγ Results Treatment of WT9-12 cells with rosiglitazone resulted in a dose-dependent and time-dependent strong inhibition of cell proliferation, an accumulation of cells in the G0/G1 phase (rosiglitazone 50 μmol/L 65.43%,rosiglitazone 100 μmol/L 64.02%, control 49.65% ) and 6% apoptosis at high concentration (rosiglitazone 200 μmol/L). Rosiglitazone reduced the phosphorylation of p70S6K in a dosedependent and time-dependent manner. The levels of phosphorylated mTOR and 4E-Bp1, the latter being a downstream substrate of mTOR related mRNA translation initiation, were not changed by rosiglitazone. Cells were pre-incubated with GW9662, a PPARγ antagonist, before the treatment with rosiglitazone, the inhibition of p70S6 kinase phosphorylation by rosiglitazone was partially prevented by GW9662 (P＜0.01). Then PPARγ siRNA was transfected into WT9-12 cells, in contrast to untransfected control or cells transfected with an irrelevant siRNA, rosiglitazone did not cause an obvious inhibition of p70S6 kinase phosphorylation in PPARγ knock-down.Conclusion Rosiglitazone inhibits the proliferation of ADPKD cystic lining epithelial cells, and down-regulates p70S6 kinase phosphorylation through mTOR-independent and PPARγ-dependent signal pathway.

Carcinoma, Renal Cell ; immunology ; pathology ; surgery ; Humans ; Keratin-19 ; metabolism ; Kidney Neoplasms ; immunology ; pathology ; surgery ; Male ; Middle Aged ; Neoplastic Cells, Circulating ; pathology ; Nephrectomy ; Vena Cava, Inferior ; pathology ; surgery

BackgroundRetinal pigment epithelial(RPE) cells can secrete platelet-derived growth factor (PDGF) and PDGF receptor(PDGFR).Studies have shown that PDGF plays a key role in the formation of proliferative vitreous retinopathy(PVR). ObjectiveThis study was to investigate the proliferation and apoptosis changes of RPE after blockage of the PDGFR-α expression by antisense oligonucleotide ( ASODN ) in vitro. Methods Human RPE cells strain was cultured in low glucose DMEM with 10％ fetal bovine serum.Logarithmic phase cells were collected and incubated in 96-well plate at the density of 5 × 105 cells/hole.PDGFR-α ASODN was transfected into RPE cells at different concentrations for 48 hours.The cells of the blank control group were regularly cultured without any transfection.The changes of PDGFR-α expression were detected by reverse transcription-polymerase chain reaction(RT-PCR),and the proliferation of RPE was detected by MTT as the A490 value.Hoechst 33258 fluorescence staining was used to determine the apoptosis of RPE.Flow cytometry method (FCM) was applied to detect the change of cell cycle and apoptosis rate of RPE cells. ResultsThe A490 values of RPE cells were 1.45±0.12,1.07±0.06,0.65±0.05 in blank control group,1.0 μmol/L Lipo-ASODN group and 2.0 μmol/L Lipo-ASODN group with the significant difference(P=0.00 ),and that of 1.0 μmol/L Lipo-ASODN group and 2.0 μ mol/L Lipo-ASODN group were significantly lower than the blank control group ( P =0.00,0.00).Hoechst 33258 staining showed that the apoptosis cells were obviously more in Lipo-ASODN group compared with blank control group.PDGFR-α ASODN transfection induced an increase of percentage of RPE cells in G0/G1 phase( F =206.70,P =0.00),and the apoptosis rates in 1.0 μmol/L Lipo-ASODN group and 2.0 μmol/L Lipo-ASODN group were significantly enhanced in comparison with blank control group ( 37.8 ± 1.3 vs 10.5 ± 0.1,61.2 ± 1.9 vs 10.5 ± 0.1 ) ( F =1808.90,P =0.00 ).Expression intensity of PDGFR-α mRNA in RPE cells in Lipo-ASODN groups was lower. ConclusionsBlocking the PDGFR-α expression with ASODN technology can suppress proliferation and induce apoptosis of RPE cells.Intensity of PDGFR-α mRNA expression in RPE cells is ASODN dose-dependent.ASODN targeted to PDGFR-α offers an experimental basis of the gene therapy for PVR.

Objective To observe the clinical efficacy of acupoint thread embedding in preventing urinary retention after cervical cancer surgery.Method Fifty eligible patients were randomized into a treatment group and a control group, 25 cases each. The control group was given catheterization after gynecological surgery; the treatment group was intervened by thread embedding at Zusanli (ST36), Sanyinjiao (SP6), Yinlingquan (SP9), Guanyuan (CV4), and Shenshu (BL23) on the 7th day after the surgery in addition to the basic intervention given to the control group. The indwelling catheter was removed on the 14th day after the surgery, and the bladder function, urination, and residual urine volume were compared between the two groups.Result After the treatment, the bladder function in the treatment group was significantly better than that in the control group (P<0.05); the markedly effective rate of spontaneous micturition was 92.0% in the treatment group versus 72.0% in the control group, and the difference was statistically significant (P<0.05).Conclusion Early intervention with acupoint thread embedding after cervical cancer surgery can effectively decrease the incidence of post-surgery urinary retention, and it's easy-to-operation and worth application in clinic.

A new species of genus shewarella Shewanellade decolorations S12, was isolated from activated sludge of a textile-printing waste-water treatment plant. In the anaerobic condition, S12 could conserve energy for growth by using Fe3 + as the terminal electron acceptor. At the optimal condition of pH8, temperature 30℃, ferric citrate 800mg/L, sodium lactate 2g/ L, yeast extract 0. 5g/ L , the cell growth increased with the raise of the amount of the Fe3+ reduction in 8k The effect of different carbon soucres, nitrogen sources, pH values and growth temperatures on the anaerobic Fe3 + reduction of Shewanella decolorations S12 was investigated. LB was favorable for Fe3 + reduction. Glucose and sodium lactate also were favorable for Fe3+ reduction. The cell growth and Fe3 + reduction increased with the raise of the amount of the yeast extract from 0 to 4g/L The amounts of the sodium lactate of 6g/ L and ferric citrate of 800mg/L were suitable for strain S12 growth and Fe3+ reduction. In the optimum initial pH value range of 6 -8 for Fe3+ reduction, strain S12 growth increased with the raise of the pH val- ue. Strain S12 could growth and reduce Fe3+ at the temperature range of 20 -40℃. The best temperature for strain S12 growth and Fe3 + reduction was 301.

OBJECTIVETo evaluate the effect of chewing sugar-free gum after sucrose challenge on dental plaque pH in situ.

METHODS16 healthy volunteers aged 23 - 32 years were screened as subjects. The pH of 48-hour dental plaque was measured using a Beetrode pH microelectrode when subjects chewed Extra sugar-free gum after sucrose challenge.

RESULTSDental plaque pH maintained at resting plaque pH when immediately chewed sugar-free gum after sucrose challenge. Chewing sugar-free gum at 5 min after sucrose challenge, dental plaque pH was raised from 5.59 (measured at 5 min after sucrose challenge) to 6.98 (measured at 10 min after sucrose challenge).

CONCLUSIONSChewing sugar-free gum after sucrose challenge can neutralize organic acid produced by bacteria in dental plaque and rapidly rise plaque pH.

Adult ; Chewing Gum ; Dental Plaque ; metabolism ; prevention & control ; Female ; Humans ; Hydrogen-Ion Concentration ; Male ; Sucrose

OBJECTIVETo study the sleep time and the prevalence of sleep disorders in children at ages of 2-12 years in Changsha City.

METHODSA total of 3756 children at ages of 2-12 years were randomly sampled from five districts of Changsha City from June 2006 to April 2007. A questionnaire survey was performed on their parents.

RESULTSThe average daily sleep time in the subjects was 10.60 hrs. The average daily sleep time among different age groups (1 year as a group separation) was different. It was 12.26, 11.57, 11.33, 11.26, 10.95, 10.64, 10.62, 10.45, 10.28, 9.83 and 9.61 hrs respectively in the 11 age groups of 2 to 12 years of age. The prevalence of sleep disorders in children at ages of 2-12 years was 40.9%, including frequent sleep snoring (8.2%), choke/gargling (1.5%), sleep apnea (0.8 %), sleep inquietude (7.6%), mouth breathing (4.9%), hyperhidrosis (22.6%), limbs spasm (3.2 %), sleep teeth grinding (9.5 %), sleep talking (5.5 %), sleep walking (0.9 %), nocturnal enuresis (2.5%), waking up by choke (1.9%), remaining wakefulness in the night due to too much daytime sleep time (1.5%), going to sleep too early (2.1%), night awakenings (1.6%), and screeching or crying during sleep (1.8%). The prevalence of different sleep disorders was different in children between boys and girls and among different age groups.

CONCLUSIONSThe average sleep time in children at ages of 2-5 years is less than the reference value recommended by the domestic child health care textbook. There is a higher prevalence rate of sleep disorders in children at ages of 2-12 years in Changsha City than the reported data in other cities.

Adolescent ; Age Factors ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Male ; Prevalence ; Sleep Wake Disorders ; epidemiology

AIM To establish an HPLC method for the content determination of six constituents in Shiyifang Vinum (Notoginseng Radix et Rhizoma,Dipsaci Radix,Carthami Flos,etc.).METHODS The content determination of notoginsenoside R1,ginsenoside Rg1,ginsenoside Rb1 and asperosaponin Ⅵ was performed on a 30℃ thermostatic Inertsil(R) ODS-3 C18column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of acetonitrile-0.1％ phosphoric acid flowing at 1.0 mL/min in a gradient manner,and the detection wavelength was set at 203 nm.The content determination of brucine and strychnine was conducted on a 30 ℃ thermostatic Geminni(R) C18 110(A) column (4.6 mm × 250 mm,5 μm),with the mobile phase comprising of acetonitrile-mixed solution of 0.01 mol/L sodium heptanesulfonate and 0.02 mol/L potassium dihydrogen phosphate flowing at 1.0 mL/min in an isocratic elution manner,and the detection wavelength was set at 260 nm.RESULTS Six constituents showed good linear relationships within their own ranges (r ＞ 0.999 0),whose average recoveries were 98.52％-99.96％ with the RSDs of 2.0％-2.3％.CONCLUSION This simple,accurate and reproducible method can be used for the quality control of Shiyifang Vinum.

OBJECTIVEHuman herpesvirus 6 (HHV-6) isolates are classified into two variants, HHV-6A and HHV-6B, based on distinct genetic, antigenic and biological characteristics. HHV-6 has been associated with encephalitis in children recently. This study aimed to establish a real time PCR assay for simultaneous detection of the two subtypes of HHV-6, and apply this new assay to children with suspected encephalitis, then analyze the relationship between the infection with HHV-6 and encephalitis in children.

METHODThe universal primers and variant-specific TaqMan probes were designed based on the highly conserved sequences of the DNA polymerase gene (U38) of HHV-6. The 5' end of the probes for HHV-6A and HHV-6B was labeled with the fluorescein reporter tetrachloro-6-carboxyfluorescein and 6-carboxyfluorescein (6-FAM), separately, while the 3' end were quenched with 6-carboxy-tetramethylrhodamine. The real time PCR assay for simultaneous detection of HHV-6A and HHV-6B was established. Then, the plasmids of HHV-6A and -6B which were diluted by a 10-fold series from 10(9) to 10(0) copies/microl, together with controls were used for testing both sensitivity and specificity of the real time PCR assay. The cerebrospinal fluid (CSF) specimens from 445 cases of suspected encephalitis were tested with this real time PCR and positive samples were then sequenced.

RESULTBoth HHV-6A (strain ZJ-159) and HHV-6B (strain GS) were positive on the real time PCR assay. There were no cross-reaction with herpes simplex virus type 1, type 2 (HSV-1, HSV-2), varicella-zoster virus (VZV), cytomegalovirus (CMV), Epstein-Barr virus (EBV), hepatitis B virus, Staphylococcus aureus, Mycoplasma pneumoniae and human DNA. A linear regression curve was obtained when plotting Ct values against the log10 of the viral DNA input for both subtypes of HHV-6. The sensitivity threshold was 10 copies/microl for the real time PCR. HHV-6 positive rate by the real time PCR assay was 4.72% (21/445), including 4 cases with HHV-6A infection, 16 cases of HHV-6B infection and 1 case with mixed HHV-6A and HHV-6B infection. The new PCR assay usually took 2 to 3 hours to provide results.

CONCLUSIONThis new real time PCR assay can simultaneously detect both subtypes of HHV-6, and have high specificity and sensitivity. It will provide an early and sensitive diagnosis of HHV-6 encephalitis in children.

Adolescent ; Child ; Child, Preschool ; DNA Fingerprinting ; DNA Primers ; DNA, Viral ; Encephalitis, Viral ; cerebrospinal fluid ; diagnosis ; virology ; Female ; Fluorometry ; Genotype ; Herpesvirus 6, Human ; genetics ; isolation & purification ; Humans ; Infant ; Infant, Newborn ; Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

Adolescent ; Biopsy ; Cystic Adenomatoid Malformation of Lung, Congenital ; diagnosis ; diagnostic imaging ; therapy ; Diagnosis, Differential ; Diagnostic Errors ; adverse effects ; prevention & control ; Humans ; Infant ; Lung Diseases ; diagnosis ; diagnostic imaging ; therapy ; Male ; Middle Lobe Syndrome ; diagnosis ; diagnostic imaging ; surgery ; Tomography, X-Ray Computed ; Treatment Outcome