Objective To investigate the expression changes of connexin 43 (Cx43) gene and the functional changes of the gap junction intercellular communication (GJIC) among cultured unstable detrusor cells and their roles in the development of detrusor instability (DI). Methods Forty healthy female Wistar rats were divided into two groups: DI group and normal control group. RT-PCR and Western blot techniques were employed to detect the expression of Cx 43 mRNA and protein in the cultured detrusor cells. Scrape loading dye transfer (SLDT) technique was used to monitor the GJIC among cultured detrusor cells. Results The expression of Cx43 mRNA and protein in DI group was much higher than that in normal control group (P

Objective To investigate the chemosensitivity of ovarian cancer SKOV3ip1 multicellular aggregates to cisplatin and taxol and to explore the possible mechanisms accounting for the effect Methods Liquid overlay system was employed to obtain multicellular aggregates (MCA) We detected the resistance with trypan blue exclusion testing, clonogenic assay, cell cycle profiles and apoptosis with flow cytometry Results MCA cells showed higher cell viability than monolayer cells ( P =0 045 and P =0 003, respectively). After 40 ?mol/L cisplatin exposure for 12 hours, no clone (≥50 cells) was formed After 10 ?mol/L taxol exposure for 12 hours, the clone formation showed significant difference in 100 cell group between multicellular aggregates and monolayer cells ( P

Objective:To investigate the mechanism of the changes of the intestinal mucosal barrier in Severe acute pancreatitis.Methods:Twenty patients with Severe acute pancreatitis were admitted and 10 volunteers as the control group.The levels of serum tumor necrosis factor alpha(TNF_?)、nitric oxide(NO)、diamine oxidase(DAO) and the concentration of plasma endotoxin(ET) were measured.The ratio of lactulose to mannitol in urine was detected by HPLC with Pulsed Electrochemical Detection.Results:Compared with the control group,significantly increasing parameters could be seen in all the patients with Severe acute pancreatitis,including the ratio of lactulose to mannitol in urine (P

Objective To evaluate the effect of low-molecular weight heparin (LMWH) on biological behavior of a human cutaneous squamous cell carcinoma cell line A431.Methods To optimize the concentration of LMWH,A431 cells were treated with different concentrations (12.5,25,50,100 and 200 IU/ml) of LMWH for 24,48 and 72 hours followed by CCK-8 assay for the detection of cell viability.Then,A431 cells were cultured with or without the presence of LMWH at 200 IU/ml for 24,48 and 72 hours.Subsequently,flow cytometry was performed to assess cell cycle,real time quantitative PCR (RT-qPCR) and Western blot to quantify the expression of vascular endothelial growth factor (VEGF) mRNA and protein respectively,double-antibody sandwich enzyme linked immunosorbent assay (ELISA) to determine the expression level of VEGF protein in the supernatant of A431 cells,wound-healing assay,Transwell assay,and adhesion assay to observe the migration and adhesivity of A431 cells.Analysis of variance and t test were carried out for statistical analysis.Results The optimal concentration of LMWH was determined as 200 IU/ml according to the CCK-8 assay,and used in the following experiment.The LMWH of 200 IU/ml resulted in a decrease in cell viability,cell cycle arrest,an increase in cell percentage in G1 phase,and a reduction in cell percentage in S phase.The proliferation index was 23.41 ± 5.51 and 11.76 ± 5.13 respectively in A431 cells at 48 and 72 hours after treatment with LMWH of 200 IU/ml,significantly lower than that in untreated A431 cells (48.62 ± 4.50,t =6.14,P ＜ 0.05; 46.86 ± 3.51,t =9.78,P ＜ 0.05).A significant decrease was observed in LMWH-treated A431 cells at 48 and 72 hours compared with the untreated A431 cells in the expression level of VEGF mRNA (10.16 ±0.07 vs.18.77 ± 0.11,4.11 ± 0.01 vs.17.39 ±0.05,t=114.38,451.10,both P＜ 0.05),VEGF protein (0.16 ± 0.01 vs.0.20 ± 0.01,0.12 ± 0.01 vs.0.21 ± 0.01,t =4.90,11.02,both P ＜ 0.05),and in the supernatant level of VEGF protein ((67.17 ± 3.34) ng/L vs. ( 122.63 ± 23.17) ng/L, (28.14 ± 3.14) ng/L vs.(86.76 ± 1.18) ng/L,t =4.10,30.27,both P＜ 0.05).The percentage of adherent cells was 29.7％ ± 1.92％ and 17.5％± 0.79％ in LMWH-treated A431 cells at 48 and 72 hours,respectively,significantly lower than that in untreated A431 cells (36.9％ ± 0.35％,34.6％ ± 0.96％,respectively,both P＜ 0.05).The migration of A431 cells was also obviously inhibited by the treatment with LMWH for 24,48 and 72 hours.Conclusion LMWH may suppress the proliferation,migration and adhesion of A431 cells via downregulating cellular viability and VEGF expression.

Objective To observe the effect of short hairpin RNA (shRNA)-mediated vascular endothelial growth factor (VEGF) gene silencing on the growth of human skin squamous cell carcinoma(SCC) xenografts in nude mice.Methods Two eukaryotic expression plasmids targeting VEGF gene,including psilencer-VEGF1-shRNA (VEGF-s1) and psilencer-VEGF2-shRNA (VEGF-s2),as well as one negative control plasmid containing random target sequence (psilencer-Target-off-shRNA,T-off),were chemically synthesized,and transfected into a human skin SCC cell line A431 to develop stably transfected cell lines.Real time quantitative PCR (RT-qPCR) and double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) were carried out to measure the expression of VEGF mRNA and protein respectively in A431 cells.Twelve nude mice were divided into 4 goups to be subcutaneously inoculated in the axillary region with untransfected A431 cells as well as A431 cells transfected with VEGF-s1,VEGF-s2 and T-off,respectively.The tumor growth was observed in nude mice every 5 days.Twenty days after the inoculation,the mice were sacrificed,and transplanted tumors were obtained from the mice and subjected to an immunohistochemical study for the measurement of VEGF,proliferating cell nuclear antigen (PCNA) and CD34 expression.Data were statistically analyzed by using the Stata 7.0 software,and t test was conducted to compare the differences between groups.Results The mRNA and protein expression levels of VEGF were significantly lower in A431 cells transfected with VEGF-s1 and VEGF-s2 than in untransfected A431 cells (27.85 ± 3.95 and 24.69 ± 2.83 vs.54.06 ± 6.38,t =6.05,7.29,both P＜ 0.01; 32.67 ± 2.52 and 29.27 ± 1.10 vs.52.85 ± 2.23,t =8.04 and 11.53,both P＜0.01 ).Twenty davs after the inoculation,the volume and weight of xenografted tumors in mice inoculated with VEGF-sl- and VEGF-s2-transfected A431 cells were significantly lower than those in mice with untransfected A431 cells ( ( 192.50 ± 10.90) mm3 and (203.67 ± 3.21 )mm3 vs.(272.00 ± 21.07) mm3,t =5.80 and 5.55,both P＜ 0.01; (0.05 ± 0.03) g and (0.13 ± 0.04) g vs.(0.25 ± 0.02) g,t =9.60 and 4.64,both P＜ 0.01 ).Decreased expression rate of VEGF,PCNA and number of CD34-positive vessels were observed in the xenografted tumor tissue from mice inoculated with VEGF-sl- and VEGF-s2-transfected A431 cells compared with that from mice with untransfected A431 cells (52.00％ ± 2.00％ and 56.67％ ± 3.06％ vs.70.00％ ± 2.00％,both P ＜ 0.01;37.01％ ± 2.41％ and 33.94％ ± 3.25％ vs.72.11％ ± 3.02％,both P＜ 0.01; 2.05 ± 0.07 and 1.72 ± 0.10 vs.4.01± 1.27,both P ＜ 0.01).No significant differences were observed in the above parameters between cells transfected with VEGF-s1- and VEGF-s2-transfected A431 cells,between untransfected and T-off-transfected A431 cells,between tumor xenografts derived from VEGF-sl- and VEGF-s2-transfected A431 cells,or between tumor xenografts derived from untransfected and T-off-transfected A431 cells (all P ＞ 0.05).Conclusions The shRNA targeting VEGF gene can significantly inhibit the expression of VEGF in A431 cells and A431-derived tumor xenografts in nude mice,in turn suppress the growth and attenuate the malignant phenotype of tumor.

Objective To provide an comprehensive evaluation of the correlation between sleep apnea hypopnea syn?drome (SAHS) and nonalcoholic fatty liver disease (NAFLD). Methods The various case-control studies on the relation?ship between SAHS and NAFLD were retrieved from all kinds of large-scale databases at home and abroad (including Web of science, EMbase, Pubmed, Springer Link, EBSCO Databases, CNKI, CQVIP, Wanfang Data). The quality evaluation of in?cluded studies was made by two independent researchers. RevMan 5.1 and stata 12.0 software were used for meta-analysis. Results A total of 11 qualified documents were included in this study. Meta analysis showed that the relative risk of NAFLD was increased in SAHS patients than non-SAHS patients (RR=2.82, 95%CI:2.03-3.92, P<0.01). The serum ala?nine aminotransferase (ALT) increased in SAHS patients (SMD=0.53, 95% CI:0.02-1.05, P < 0.05). Compared with non-SAHS patients, the apnea-hypopnea index (AHI) was significantly higher in SAHS patients combined with severe NAFLD than those combined with mild NAFLD (SMD=1.42, 95%CI:0.12-2.72, P < 0.05). Conclusion The risk of NAFLD in?creases in SAHS patients. The severity of NAFLD is relatively higher with the severity of intermittent hypoxia.

Objective To investigate the role of bladder interstitial cells of Cajal(ICCs) in regulating the excitability of detrusor myocytes.Methods 1)Ultrastructural connection between ICCs and detrusor myocytes was detected by transmission electron microscope(TEM).2) Fluorescence redistribution after photobleaching(FRAP) was carried out to detect the intercellular communication between detrusor myocytes and its neighboring ICCs.Results 1)Gap junction between the 2 kinds of cells was confirmed by TEM.2)After target detrusor myocyte was photobleached,fluorescence intensity in detrusor myocytes was recovered gradually.In the meantime,fluorescence intensity in the ICCs neighboring to the target detrusor myocyte was decreased.This indicates a signal transmission from the neighboring ICCs to the target detrusor myocyte.Conclusion Structural connection is seen between ICCs and detrusor myocytes.Excitability signals might be transferred from ICCs to detrusor myocyte.

Objective: To investigate the effect of survivin and bcl-2 antisense oligodeoxynucleotides ( AsODN) combined transfection on the human gallbladder carcinoma cell line GBC-SD in vitro. Methods: Survivin and Bcl-2 protein expressions were detected by immunohistochemical method; Cultured cells were divided into 4 groups: Nomal control group, survivin antisense observed group, bcl-2 antisense observed group and combined group. After transfected for 24 h, the expression of survivin mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR). Cell morphological changes were observed under electron microscopy. Apoptosis index (AI) was examined by flow cytometry; Inhibitory rate (IR) was determined by the colorimetri MTT cell viability and proliferation assay. Results: Survivin and Bcl-2 protein were highly expressed in gall bladder carcinoma cells; The expression of survivin mRNA was decreased 47. 8%. Abnormal morphological changes of cells were observed in the three AsODN transfection groups; The AI in survivin antisense observed group,bcl-2 antisense observed group,and combined group was 11.38%?3.91% , 9.26%?4.15% , 28.45%?6.34% respectively and significantly higher than the nomal control group (P

Objective To study the apoptosis of gallbladder carcinoma cell line GBC-SD induced by antisense oligodeoxynucleotide (ASODN) targeting survivin. Methods ASODN targeting survivin was transfected into GBC-SD cells mediated by lipofectin. Cultured cells were divided into 3 groups: control group,sense oligonucleotide (SODN) group and ASODN group. After transfected for 16 h, the cultured cells were harvested and the following texts were carried out. The expression of survivin mRNA was detected by RT-PCR. Flow cytometer were used to detect apoptosis. Morphological changes were observed by electron microscopy. Results The expression of survivin mRNA was decreased 47.83% in ASODN group while apoptosis was increased from (0.50?0.23)% to (26.28? 3.91)%. Abnormal morphological changes of cells were observed in ASODN group and apoptosis bodies were found in some gallbladder carcinoma cells. Conclusion The expression of survivin may be decreased in GBC-SD cells after ASODN transfection.ASODN targeting survivin could induce gallbladder carcinoma cells apoptosis effectively.

Ascoviruses, iridoviruses, asfarviruses and poxviruses are all cytoplasmic DNA viruses. The evolutionary origins of cytoplasmic DNA viruses have never been fully addressed. Morphological, genetic and molecular data were used to test if all four cytoplasmic virus families (Ascoviridae, Iridoviridae, Asfarviridae, and Poxvirirdae) evolved from nuclear replicating baculoviruses and how the four virus groups are related. Molecular phylogenetic analyses using DNA polymerase predicted that cytoplasmic DNA viruses might have evolved from nuclear replicating baculoviruses, and that poxviruses and asfarviruses share a common ancestor with iridoviruses. These three cytoplasmic viruses again shared a common ancestor with ascoviruses. Morphological and genetic data predicted the same evolutionary trend as molecular data predicted. A genome sequence comparison showed that ascoviruses have more baculovirus protein homologues than do iridoviruses, which suggested that ascoviruses have evolved from baculoviruses and iridoviruses evolved from ascoviruses. Poxviruses showed genetic and morphological similarity to other cytoplamic viruses, such as ascoviruses, suggesting it has undergone reticulate evolution via hybridization, recombination and lateral gene transfer with other viruses. Within the ascovirus family, we tested if molecular phylogenetic analyses agree with biological inference; that is, ascovirus had an evolutionary trend of increasing genome size, expanding host range and widening tissue tropism for these viruses. Both molecular and biological data predicted this evolutionary trend. The phylogenetic relationship among the four species of ascovirus was predicted to be that TnAV-2 and HvAV-3 shared a common ancestor with SfAV-1 and the three virus species again shared a common ancestor with DpAV-4.