LncRNA HULC promots HCC growth by downregulating miR-29 / 中华肿瘤杂志

Objective@#To explore the effects of lncRNA HULC on hepatocellular carcinoma (HCC) growth by down-regulating miR-29.@*Methods@#The expression levels of HULC and miR-29 in HCC tissues and cells were detected by real-time quantitative PCR (RT-qPCR), and the correlation analysis was performed. After HCC cells were transfected with HULC overexpressed plasmid or siRNA, the expressions of miR-29 and its target gene SETDB1 were determinate by RT-qPCR. According to the bioinformatic prediction of the miR-29 binding site in the HULC sequence, the report gene plasmids were constructed. The HCC cells were co-transfected with miR-29 mimics or miR-29 inhibitor, and the HULC targeted regulation of miR-29 was verified by dual luciferase reporter assay. The effect of miR-29 on the HULC-mediated proliferation in HCC cells was detected by cell count kit 8 (CCK-8) experiment. Expression of tumor proliferation antigen Ki-67 was detected by RT-qPCR.The Hep3B cells were inoculated in mice and miR-29 mimics and miR-29 negative control (NC) further injected into the lesions. The tumor volume was observed, and the expressions of tumor proliferation antigen ki-67 in tumor tissues were detected by immunohistochemical staining.@*Results@#The expression of HULC was significantly up-regulated while the expression of miR-29 was significantly down-regulated in HCC tissues and cells (P<0.01). The level of HULC was negatively correlated with miR-29 in tumor tissues (r=-0.754, P<0.01) and HCC cells (r=-0.865, P<0.05). The in vitro experiments showed that, compared with the blank control group, the expression of miR-29 in HULC overexpressed Huh7 cells was significantly reduced, while the mRNA level of miR-29 target gene SETDB1 was increased (P<0.01). The expression of miR-29 was significantly increased in HULC deleted Hep3B cells, while the mRNA expression of SETDB1 was decreased (P<0.01). Double luciferase reporter gene assay showed that miR-29 mimics significantly inhibited the luciferase activity of Hep3B cells transfected with HULC wide type (psi-HULC-WT) plasmid but had no effect on Hep3B cells transfected with mutant plasmid (psi-HULC-Mut). However, the miR-29 inhibitor antagonized the inhibitory effect of miR-29 mimics on luciferase activity of psi-HULC-WT (P<0.01). Cell proliferation experiments showed that, compared with the control group, the proliferation ability of miR-29 mimics overexpressed Huh7 cells was significantly reduced.After 24, 48 and 72 hours of treatment, the proliferation rates of Huh7 cells in the HULC overexpressed group were (43.87±3.82)%, (83.45±7.46)% and (123.34±8.67)%, respectively, significantly higher than (13.45±1.77)%, (23.54±1.37)% and (38.21±2.09)% of control group (P<0.05). After treatment for 48 and 72 hours, the proliferation rates of miR-29 mimics transfected Huh7 cells were (57.10±1.94)% and (73.76±3.46)%, respectively, significantly lower than (83.45±7.46)% and (123.34±8.67)% of control group (P<0.05). After treatment for 48 and 72 hours, the proliferation rates of Huh7 cells transfected with miR-29 mimics and miR-29 inhibitor group were (76.45±3.24)% and (89.37±4.37)%, respectively, significant higher than (57.10%±1.94)% and (73.76±3.46)% of the control group (P<0.05). After 48 h transfection, the expression of Ki-67 in Huh7 transfected with miR-29 mimics was significantly inhibited compared with the control group (P<0.01). However, the expression of Ki-67 mRNA was increased in Huh7 cells transfected with miR-29 inhibitor (P<0.01). The results of in vivo experiments showed that the tumor volumes of the control group, miR-29 mimics group and miR-29 mimics + miR-29 inhibitors group were (504.0±19.6) mm3, (310.0±24.3) mm3 and (483.7±21.2) mm3, respectively. Injection of miR-29 mimics reduced while miR-29 inhibitor promoted tumorigenesis ability of Huh7 in nude mice (P<0.01). The immunohistochemical staining showed that the average optical density values of Ki-67 protein in tumor tissues of the control group, miR-29 mimics group and miR-29 analogue+ miR-29 inhibitor group were 0.65±0.08, 0.36±0.07 and 0.56±0.06, respectively. The expression level of Ki-67 protein in miR-29 mimics group was significantly reduced (P<0.01) while increased in the miR-29 mimics+ miR-29 inhibitor group (P<0.01).@*Conclusion@#LncRNA HULC promotes HCC growth by down-regulating miR-29.

Preparation of VEGF-loaded PLGA fluorescent sub-micrometer spheres and in vitro controlled release characteristics / 重庆医学

Objective To prepare the biodegradable polylactic acid glycolic acid (PLGA) copolymer.encapsulated vascular endothelial growth factor(VEGF) loaded fluorescent controlled release sub-microspheres,to understand the efficiency of microspheres loading and releasing VEGF and to observe in vitro microspheres degradation.Methods VEGF-loading PLGA sub-microspheres were prepared by the two-phase solvent evaporation method,the in vitro degradation of fluorescent microspheres was observed by the laser confocal scanning electron microscopy.The drug loading efficiency and drug release curve were observed by ELISA.Results The VEGF loading PLGA fluorescent microspheres were successfully prepared by using the two-phase solvent evaporation method.The microspheres morphology was normal by using the scanning electron microscope and laser confocal microscope.The particle size was 0.5-1.0 μm with the laser particle size analyzer.The distribution was homogeneous.The VEGF loading rate and encapsulation rate detected by the quantitative ELISA were 3.91％ and 51.42 ％ respectively.The fluorescent microscope observed their slow degradation.The VEGF gentle release was detected by the quantitative ELISA,which showing linear zero order release trend.Conclusion The drug loading efficiency of VEGF-loading PLGA microspheres with 0.5-1.0 μm diameter is higher with linear zero order release,which can be directly observed by fluorescent light.

Correlation of C - erbB - 2 Status and Histologic Features in 1 6 3 Invasive Breast Carcinomas of baoji area / 中国基层医药

ObjectiveTo study the relationship between C-erbB-2 and estrogen (ER) and progesterone (PR) receptors, and the relationship between C-erbB-2, ER, PR with histologic grade. MethodsTo detect ER, PR and C-erbB-2 states by using immunohistochemical analysis and fluorescence in situ hybridization for C-erbB-2 in 163 unselected invasive breast carcinomas. ResultsC-erbB-2, ER ,PR were expressed in 21.5％ ,64.4％ ,44.2％ of 163 cases respectivly . 5 pure mucinous carcinomas , 3 tubular carcinomas and 1 micropapillary carcinoma were ER + ( 100.0％ ) 、C-erbB-2 - ( 100.0％ ) and PR + (40.0％ ,66.7％, 100.0％ ). C-erbB-2 was positive in 22.3％ of grade Ⅱ and 27.0％ of grade Ⅲ invasive ductal carcinomas and negative in all grade Ⅰ invasive ductal carcinomas.ER and PR expression were decreased significantly in C-erbB-2 + tumors compared with C-erbB-2 - tumors( ER,25. 7％ vs 75.0％ ; PR,25.7％ vs 49.2％ ). Although ER or PR expression is decreased in C-erbB-2 + tumors, a substantial proportion of them still express ER or PR. ConclusionC-erbB-2 overexpression or amplifcation was limited to a minority of invasive breast carcinomas. Tumour grade was an independent predictor for ER expression. ER was expressed in small number of high-grade and in large number of grade Ⅰ invasive ductal carcinomas. C-erbB-2 overexpression or amplification essentially was limited to grades Ⅱ and Ⅲ ductal carcinomas and correlated inversely with ER or PR expression.

Expression of connexin 43 gene and the gap junction intercellular communication among cultured unstable detrusor cells / 第三军医大学学报

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

Interstitial cells of Cajal regulates excitability of detrusor myocytes: structure evidence / 第三军医大学学报

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.