Objective To explore the effect of estradiol benzoate on the expressions of heat shock protein (HSP)70, 90 in endometrial glandular epithelial cell. Methods Normal endometrial glandular epithelial cells were isolated, and cultured by enzymolysis method and identified by electron microscopy and immunohistochemical analysis. The normal endometrial glandular epithelial cells were treated with culture medium only, estradiol benzoate (10 -9 , 10 -8 , 10 -7 or 10 -6 mol/L), estradiol benzoate(10 -9 , 10 -8 , 10 -7 or 10 -6 mol/L) and antiestrogen ICI 182780(fulvestrant,faslodex, 1?mol/L)and ICI 182780 only for 6, 12, 18, 24 hours respectively. The dose-and time-related effect of estradiol benzoate and ICI 182780 on the cell growth was measured by mononuclear cell direct cytotoxicity assay (methyl thiazolyl tetrazolium assay), and that on the expression of HSP70 and HSP90 in normal endometrial glandular epithelial cell in vitro was measured by immunohistochemical analysis and computerized image analysis system. Results Estradiol benzoate stimulated cell growth in a time-and dose-dependent manner and the effect was attenuated by the antiestrogen ICI 182780. The average cell growth rates of 10 -9 , 10 -8 , 10 -7 , 10 -6 mol/L estradiol benzoate for 24 hours were(170?9)%,(207?11)%,(231?12)%,(257?10)%, which were significantly higher than those of 6 hours (117?13)%, (129?10)%, (146?10)%, (176?6)%, P

AIM: To construct the replicative defecient adenovirus Ad-Runx3 expressing Runx3, and to express it in U251 malignant glioblastoma cells. METHODS: The runx3 gene with a flag tag was amplified by PCR using pCMV5-AML2 as a template, and was confirmed by DNA sequencing. The adenovirus shuttle vector pShuttle-CMVRunx3 was constructed by introducing runx3 DNA fragment into the sites of Kpn Ⅰ and Xho Ⅰ of pShuttle-CMV vector. This recombinant plasmid was linearized by Pmel and electronically transfected into BJ5183 cells to get the recombinant adenovirus vector Ad-Runx3. The recombinant adenovirus expressing Runx3 was infected into U251 malignant glioblastoma cells. The expression of exogenous Runx3 was observed by immonoblotting and its localization was detected by immunostaining using anti-Flag tag antibody. RESULTS: The recombinant adenovirus expressing Runx3 with a Flag tag was constructed and infected into U251 glioblastoma cells. The exogenous Runx3 protein was detected only in the nuclei. CONCLUSION: The recombinant adenovirus expressing Runx3 with a Flag tag is constructed successfully, and the Runx3 protein expressed in the nuclei of infected cells. The study laid a foundation for further research of the function of Runx3 in gliocarcinogenesis.

Objective To investigate the effect and mechanism of miR-27a-3p on nerve cell apoptosis induced by oxygen glucose deprivation/reoxygenation(OGD/R)through regulation of Rho family GTPase 3(Rnd3)expression.Methods PC12 neurons were cultured in vitro and reoxygenated for 3,6,9 h and 12 h after 2 h oxygen glucose deprivation.Cell viability,miR-27a-3p expression and Rnd3 mRNA expression were assessed at each time point and the optimal reoxygenation time point was screened.After transfection of miR-27a-3p Mimic,miR-27a-3p Inhibitor and their negative control,transfection of shRnd3 and its negative control,or co-transfection of shRnd3 and miR-27a-3p Inhibitor through lentivirus,CCK-8 assay was used to detect cell activity.The apoptosis rate of the cells was detected using flow cytometry.Expression of miR-27a-3p and Rnd3 mRNA was detected by RT-qPCR.Expression of apoptosis-related protein and Rnd3 protein was detected by Western blot.The dual luciferase reporter assay confirmed the targeting relationship between miR-27a-3p and Rnd3.Results Upregulation of miR-27a-3p increased cell viability,decreased total cell apoptosis rate,suppressed pro-apoptotic proteins Cleaved Caspase-3(C-caspase-3)and Bax,and promoted expression of anti-apoptotic protein Bcl-2(P<0.05);The opposite result was found when down-regulating miR-27a-3p.The double luciferase reporter gene assay showed that Rnd3 was the target gene of miR-27a-3p.Down-regulation of Rnd3 increased cell viability,decreased the total rate of apoptosis,suppressed the pro-apoptotic protein C-caspase-3,Bax,and promoted expression of the anti-apoptotic protein Bcl-2(P<0.05).However,miR-27a-3p Inhibitor reversed the protective effect of shRnd3.Conclusion miR-27a-3p alleviates OGD/R-induced damage to PC12 neurons by targeting Rnd3 to inhibit cell apoptosis.

Objective To explore the feasibility and efficacy of transcatheter closure of ventricular septal defect (VSD) through radial artery combined femoral vein approach. Methods A total of 11 patients with congenital VSD, who were admitted to authors' hospital during the period from June 2017 to November2017, were enrolled in this study. The patterns of lesion included intracristal type (n=3) and perimembranous type (n=8), and in 3 patients the VSD was associated with concant ventricular septal aneurysm. Transcatheter closure of VSD via radial approach was carried out in all patients. The mean age of the patients was (37.82±12.44) years old, and the average body weight was (62.79±14.95) kg. The transthoracic echocardiography (TTE) showed that the mean diameter of VSD was (5.87±1.91)mm. The effect of transcatheter closure therapy was assessed by intraoperative TTE and left ventriculography. All patients were followed up with electrocardiogram and TTE at 24 hours and one, 3, 6 months after transcatheter closure therapy. Results Successful closure was achieved in 10 patients, and one patient had to be transferred to surgery because the catheter could not pass through the defect. The mean diameter of the implanted occluders was (7.50±3.60)mm, the average procedural time and fluoroscopy time were (47.20±5.45) min and (13.00±3.65) min respectively. The postoperative average in-bed time was (99.00±11.97) min. Two patients developed radial artery spasm during the operation. During the follow-up period lasting for a mean of (3.50±1.90) months, no serious complications, such as dropping of occluder, residual shunt, atrioventricular block, aortic regurgitation, radial artery occlusion, etc. occurred in the 10 patients. Conclusion For the treatment of VSD, transcatheter closure through radial artery combined with femoral vein approach is safe and effective. Therefore, this technique is worthy of clinical application.

Objective To investigate the effect of salidroside on intestinal mucosal immune status in rats under compound stress of hypoxia and training (HTCS) and the mechanism. Methods SD rats were randomly divided into HTCS model group (model), placebo group (placebo) and salidroside group (salidro). Model group received no intervention, and placebo and salidro group received intraperitoneal injection of normal saline and salidroside, respectively. Then, ileum tissue of rats were collected and the intestinal damage was assayed by HE staining and Chiu scores. Intestinal permeability indices, including serum D-diamine oxidase (DAO), D-lactic acid (DLA) and endotoxin (END) and secretory immunoglobulin A (sIgA) of intestinal tissue were detected by ELISA. T lymphocyte subsets of intestinal tissue were detected by flow cytometry. Expression of tight junction molecules, including ZO-1, Claudin-3, occluding, were detected by PCR and western blot. Activation of TLR4/NF-κB signaling pathway was detected by Western blot analysis. Results Compared with model group and placebo group, salidro group had the decreased intestinal mucosal injury and low Chiu score, and the level of intestinal permeability indices including serum DAO, DLA and END fell off. CD4⁺ T cell percentage, CD4⁺/CD8⁺ ratio and sIgA level were went up, while CD8⁺ T cell percentage was went down. mRNA and the level of protein expressions of ZO-1, claudin-3 and occludin increased, while activation of TLR4/NF-κB signaling pathway was inhibited. Conclusion Salidroside can alleviate the intestinal barrier injury and improve intestinal mucosal immune status of rats under compound stress of hypoxia and training via inhibiting TLR4/NF-κB signalling pathway.
Animals ; Rats ; Rats, Sprague-Dawley ; NF-kappa B ; Toll-Like Receptor 4/genetics* ; Claudin-3 ; Hypoxia ; Immunoglobulin A, Secretory ; Signal Transduction