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Background: Pancreatic cancer is among the most lethal malignancies worldwide. In this study, we aimed to determine whether miR-573 could suppress pancreatic cancer cell proliferation, migration, and invasion by targeting E2F3. Materials and Methods: MiR-573 expression in pancreatic cancer tissues and cell lines was measured using real-time PCR. Target genes of miR-573 were screened using bioinformatics tools and confirmed using dual-luciferase reporter assay and real-time PCR. Pancreatic cancer cells were transfected using an miR-573 mimic or siRNA E2F3. Furthermore, cell proliferation, migration, and invasion were assessed using CCK-8, Edu staining, colony-forming assay, wound healing assay, and transwell assay in vitro. The in vivo effects of miR-573 were verified using tumor xenografts. Differential expression and prognostic analyses of miR-573 and E2F3 were visualized using the KaplanMeier plotter and GEPIA. Results: We found that the expression of miR-573 was significantly reduced in pancreatic cancer tissues and cell lines. Overexpression of miR-573 obviously suppressed the proliferation, migration, and invasion of pancreatic cancer cells. The Dual-luciferase assay showed that miR-573 could specifically target E2F3. Furthermore, E2F3 was up-regulated in pancreatic cancer tissues and cell lines and E2F3 down-regulation inhibited the proliferation, migration, and invasion of pancreatic cancer cells. The ectopic expression of miR-573 inhibited xenograft tumor growth in vivo. Results from the Kaplan-Meier analysis and GEPIA showed that patients with a high level of miR-573 had a significantly reduced risk of death while those with a high level of E2F3 displayed significant correlation with the tumor stage and suffered worse prognosis. Conclusions: MiR-573 could suppress the proliferation, migration, and invasion of pancreatic cancer cells by targeting E2F3, thereby establishing miR-573 as a novel regulator of E2F3 and indicating its critical role in tumorigenesis, especially in pancreatic cancer.
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Objective To evaluate effect of Xugu-Huoxue decoction on the treatment of patients with femoral neck fracture after internal fixation with hollow screw fixation. Methods A total of 120 patients with femoral neck fracture were randomly divided into the treatment group and the control group according to the inclusion criteria, 60 patients in each group. The control group was treated with cannulated screw internal fixation; the observation group was treated with Xugu-Huoxue decoction at the second day after the operation. Both groups were assessed with the Harris hip function score at the end of 12 months after operation. The SF-36 scale was used to evaluate the quality of life of the patients. Results The total effect rate of the observation group was 88.3% (53/60) and the control group was 71.7% (43/60). The difference between the 2 groups was statistically significant (χ2=5.208, P=0.022). At 12 months after surgery, the pain (41.4 ± 7.3 vs. 32.2 ± 5.7, t=7.738), the joint function (45.7 ± 6.2 vs. 36.3 ± 7.2, t=7.701), joint mobility (5.0 ± 0.8 vs. 3.1 ± 0.8, t=13.115), and the total score of Harris (87.7 ± 4.6 vs. 65.4 ± 5.4, t=24.461) in the observation group were significantly higher than those in the control group (P<0.001). The physical function (83.1 ± 7.2 vs. 78.8 ±14.2, t=2.095), body pain (82.1 ± 9.9 vs. 67.7 ± 11.1, t=7.524), the overall health (76.6 ± 10.3 vs. 68.8 ± 14.4, t=3.401) and activity (81.1 ± 7.9 vs. 76.6 ± 11.2, t=2.549) in the observation group were higher than those in the control group (P<0.05 or P<0.01). Conclusions The Xugu-Huoxue decoction could improve the prognosis of patients with femoral neck fractures after cannulated screw fixation, promote the recovery of hip function and improve the quality of life.
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Objective To observe the therapeutic effect between less invasive stabilization system (LISS) plate and anatomical plate internal fixation for the treatment of tibial plateau fractures. Methods The clinical data of 58 patients with tibial plateau fractures were retrospectively analyzed. The patients were divided into LISS plate group and anatomical plate group according to the internal fixation method with 29 cases each. The operative time, blood loss, incision length, fracture healing time, postoperative weight-bearing time, postoperative complications and therapeutic effect (according to Rasmussen knee joint function score scale) were compared between 2 groups. Results The operative time, incision length, blood loss and incidence of postoperative complications in LISS plate group were significantly lower than those in anatomical plate group:(68.5 ± 7.1) min vs. (92.3 ± 9.4) min, (5.8 ± 1.4) cm vs. (8.6 ± 2.1) cm, (208.5 ± 27.8) ml vs. (329.7 ± 25.2) ml and 17.2%(5/29) vs. 41.4%(12/29), and the excellent rate was significantly higher than that in anatomical plate group: 86.2% 25/29) vs. 62.1%(18/29), and there were statistical differences (P0.05). Conclusions Both of LISS plate and anatomical plate internal fixation for the treatment of tibial plateau fractures have good clinical efficacy, but the LISS plate has advantages of shorter operation time, less tissue trauma, less blood loss, quicker fracture healing and less postoperative adverse reaction.
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Objective To investigate the influence of streptozocin (STZ)'s dose on the inductive effect of diabetes in C57BL/6J mice, and investigate the dose-effect relationship and the optimal dose range. Methods 145 C57BL/6J mice were randomly divided into 9 diabetic groups (group A to group 1, n = 15 in each group) and I control group (n = I0) to receive intraperitoneal injection of STZ with the dosages of 30, 60, 80, 100, 120, 150, 180, 210, 240 mg/kg and same amount of buffer solution,respectively. Changes of blood glucose, body weight, survival rate at 45 day and serum insulin level were monitored, and the relationship with STZ doses was analyzed. Pancreas and kidneys of the mice were removed for morphological examination, and immunohistochemistry was used for determination of insulin in pancreas and CD<,68> in kidneys. Results Compared with control group, blood glucose in group C ～G increased significantly; body weigh, insulin level decreased significantly (P < 0.05), and the STZ dose was positively correlated with mean blood glucose (r = -0.984, P < 0.05) and was negatively correlated with mean serum insulin levels (r = 0.994, P <0.05). The diabetes modeling rates in group C ～ G (86.7% ～ 100%) were higher than those of group A and B (0 and 40%, P<0.05). At the 45th day, the survival rates of group C ～G (46.7% ～ 73.3%) were higher than those of group H and 1 (13.3% and 0, P <0.05). There was no obvious injury of pancreas and kidneys in group B, whereas, in group C and G, pancreatic island atrophy and decreased insulin secretion were observed; deposits of extracellular matrix and macrophage increased in the mesangium were also present. Conclusions 80 ～ 180 mg/kg of STZ dose was ideal for establishing diabetes model in C57BL/6J mice. Within this range, the modeling rate and survival rate was higher, and target organs injury was typical. The STZ dose was positively correlated with blood glucose and negatively correlated with serum insulin levels.
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Objective To observe the effects of differentiation of mature islet cells of mice on marrowmesenchymal stem cells (mMSCs). To provide transplant source for islet cell transplantation in the treatment of diabetes. Methods The culture, isolation and passage of mMSCs was performed by using patch wall, cell shape was observed by confocal microscope, and flow cytometry analysis was used to determine their biological characteristics. The type Ⅳ collagenase was injected into common bile duct to digest the pancreas, and then gradient centrifugation was used to isolate islet cells. The transwell co-culture system was used for third generation of mMSCs and isolated islet cells, then inversion microscope was used to observe the cell growth and morphological changes, immunochemistry methods was applied to detect the expression of insulin in mMSCs, and insulin release test was performed to determine the secretion of insulin. The control group consisted of cultured mMSCs alone. Results The cells from mouse bone marrow were found to be in long spindle shape with large volume after 48 hours in culture. One week later the cells grew in the form of colony with serial subcultivation. The cell surface molecules including Sca-l, CD29, CD44, CD105 were positive with high level of expression;while the cell surface molecules including CD34, CD45 were negative, all of these results confirmed that the ceils were mMSCs. After 7 days of coculture with mice islet cells, part of mMSCs cells were positively stained by insulin immunohistochemisty, the insulin secretion was (16.83±0. 15)μIU/ml.Conclusions After cocultured with islet cells, mMSCs isolated from mouse bone marrow could differentiate into islet like cells. These cells may be used in the islet cells transplantation in the treatment of diabetes, which provided a solution to the problems of donor-shortage and immunologic rejection.
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Objective:To construct a retroviral vector carrying both enhanced green fluorescent protein(EGFP) and human insulin gene for transfecting the bone marrow mesenchymal stem cells (BMSCs),and to observe the treatment effect of transfected BMSCs after transplanted into diabetes mice.Methods:Two gene segments of insulin-IRES-EGFP and IRES-EGFP were obtained by overlap extension PCR technology,and then the 2 segments were cloned into retroviral vector (pMSCV).The vectors were used to transfect BMSCs after packaged with PT67 cells.The expression of EGFP was observed by inverted fluorescence microscope and the expression of insulin gene was examined by RT-PCR in the infected BMSCs.The transfected BMSCs with 2 viruses were transplanted into diabetic mice separately.The blood glucose levels and body weights of mice were examined in 2 groups,and the results were compared with those of normal control group.Results:The recombinant retroviral vectors pMSCV-insulin-IRES-EGFP and pMSCV-IRES-EGFP were successfully constructed.The BMSCs infected by both vector stably gave out green fluorescence under microscope; the cells infected with pMSCV-insulin-IRES-EGFP also stably expressed human insulin gene.The blood glucose level of the mice transplanted with BMSCs transfected with pMSCV-insulin-IRES-EGFP was significantly lower than that transplanted with BMSCs transfected with pMSCV-IRES-EFGP (P
