ABSTRACT
Objective:To study the function and mechanism of miR-30a in rat cerebral ischemia-reperfusion (I/R) injury.Methods:The middle cerebral artery occlusion (MCAO) model was established by intravascular suture method, and the expression of miR-30a in brain tissue was detected by real-time quantitative polymerase chain reaction (qRT-PCR). After intracerebroventricular injection of miR-30a lentivirus, the infarct area was detected by 2, 3, 5-triphenyltetrazole chloride (TTC) staining, the neurological deficit was detected by Bederson method, and the concentration of neurotrophin-3 (3-NT) and nitric oxide (NO) in brain tissue was detected by enzyme-linked immunosorbent assay (ELISA). The protein levels of kelch like ECH associated protein 1 (Keap1), NF-E2-related factor 2 (Nrf-2) and hemeoxygenase-1 (HO-1) in brain tissue were detected by Western blot. Double luciferase reporter assay was used to detect the targeting relationship between miR-30a and Keap1.Results:Compared with sham operation group, the expression of miR-30a was down-regulated in a time-dependent manner after I/R. The overexpression of miR-30a can reduce the area of cerebral infarction tissue at the pathological level, the degree of neurological impairment at the functional level, the 3-NT, NO and Keap1 at the molecular level, and enhance the expression of Nrf2 and HO-1. The dual luciferase reporter assay also showed that miR-30a could bind to Keap1 mRNA.Conclusions:The expression of miR-30a was down-regulated in MCAO rat brain tissue, and miR-30a could attenuate cerebral I/R injury in rats by targeting Keap1.
ABSTRACT
Objective To evaluate radiofrequency ablation (RFA) assisted splenic preservation in traumatic splenic rupture.Methods Data of 70 cases with traumatic rupture of the spleen at our hospital from Septembcr 2009 to June 2014 were retrospectively analysed.Patients were divided into two groups according to different surgical methods,namely,RFA group (n =35) and control conventional surgery group (n =35).Results In the RFA group,34 cases underwent successful spleen preserving operation,the success rate was 97％.In control group,26/35 cases received splenic preservation operations successfully,the success rate was 74％ (26/35) (x2 =7.467,P ＜ 0.05).Average operation time,bleeding during operation and intra-operative transfusion in RFA group were (80 ± 22) min,(116 ± 66) ml and 5 cases,rcspectively,significantly better than that in control of group.(122 ± 80) min,(237 ± 192) ml and 13 cases,respectively (t =4.58,t =3.324;x2 =4.786,P ＜ 0.05).Postoperative bleeding,hospital stay and cases receiving transfusion in RFA group were (113 ± 72)ml,(7.8 ± 1.2) d and 2 cases,respectively,which were remarkably better than those in control group of (246 ± 140) ml,(10.2 ± 1.6) d and 8 cases (t =3.267,t =4.536;x2 =4.9;P ＜ 0.05).Postoperative complications in the two groups were similar (x2 =0.913,P ＞ 0.05).Conclusions Compared with traditional spleen preserving surgery,RFA is simple and effective.It can greatly reduce the difficulty and risks of splenic preservation surgery,increasing the preservation of ruptured spleen.
ABSTRACT
Objective To investigate the effects of silencing TSG-6 gene modified bone marrow mesenchymal stem cells (BMSC) transplantation on liver allotransplantation rejection in rats.Methods BMSC and KCs were isolated from rats and cultured in complete medium.We down-regulated TSG-6 expression of BMSC with lentiviral vectors carrying short hairpin RNA (LV3-shTSG-6).TSG-6mRNA and protein level of BMSC were tested respectively by quantitative real-time PCR and western blot analysis.After co-cuhured between BMSC and KCs,the expression of TNF-α and TSG-6 in cell supernatants were tested.Then,we established the orthotopic liver transplantation models in rats,the rats of each group were killed at 1 days,3 days and 7 days after operation.The serum levels of AST,ALT,TBIL,γ-GGT,TNF-α,IL-6 and IL-4 were tested with ELISA in each group,TSG-6mRNA and protein level of liver tissues obtained from each group were tested by quantitative real-time PCR and western blot analysis.The 1iver tissues of each group were stained with HE,then microstructure of liver tissues were observed under light microscope.The postoperative survival rates in other rats of each group were observed.Results The lentivirus transfection efficiency of mesenchymal stem cells was beyond 70 percent;After co-cultured between BMSC and KCs,the expression of TSG-6 in TSG-6-shRNA-BMSC + KCs group supernatant was significantly lower in different time point than that in BMSC + KCs group and TSG-6-NC-BMSC + KCs group (P ＜0.05),and the expression of TNF-α peak in BMSC + KCs group and TSG-6-NC-BMSC + KCs group supernatants were at 6 hours,which were significandy lower than those at 12 hours (P ＜0.05),but the expression of TNF-α in TSG-6-shRNABMSC + KCs group at 12 hours was significantly higher than those in BMSC + KCs group and TSG-6-NC-BMSC +KCs group (P ＜0.05);After transplantation,the serum levels of AST,ALT,TBIL,γ-GGT,TNF-α and IL-6 in TSG-6-shRNA-BMSC group was significantly higher than those in TSG-6-NC-BMSC group and BMSC group in different time point (P ＜ 0.05),but had no significant difference compared with PBS group;The serum levels of IL-4 in TSG-6-shRNA-BMSC group,TSG-6-NC-BMSC group and BMSC group was significantly higher than that in PBS group(P ＜ 0.05),but TSG-6-shRNA-BMSC group compared with TSG-6-NC-BMSC group and BMSC group,the serum levels of IL-4 was significantly lower (P ＜ 0.05);In pathological changes,we found that the degree of liver rejection in TSG-6-shRNA-BMSC group and PBS group were seriously obvious,and were graded Ⅱ-Ⅲ with Banff schedule;Comparation of postoperative survival time in each group,TSG-6-shRNA-BMSC group and PBS group were (16.6 ±4.6) d and (15.4 ± 6.7) d respectively,which were signifcantly lower than those in TSG-6-NCBMSC group (69.6 ± 28.1) d and BMSC group (69.2 ± 28.2) d (P ＜ 0.05).Conclusion Transplantation of BMSC secreted TSG-6 could,to some extent,mitigate acute liver transplantation rejection.