MDM4 is targeted by miR-449b-5p to promote the proliferation of endometrial carcinoma.

OBJECTIVE: Previous studies have shown the involvement of microRNA-449b-5p (miR-449b-5p) and MDM4 in tumor development. This study aims to illustrate the role of miR-449b-5p in inhibiting proliferative capacity of endometrial carcinoma (EC) by targeting MDM4. PATIENTS AND METHODS: Expression levels of miR-449b-5p and MDM4 in tumor tissues and paracancerous ones of EC patients were determined. Relationships between their levels and clinical parameters of EC patients were analyzed. Subsequently, regulatory effects of miR-449b-5p and MDM4 on proliferative capacities in KLE and HEC-1B cells were assessed by cell counting kit-8 (CCK-8) and colony formation assay, respectively. Thereafter, in vivo xenograft models were established in nude mice administrated with KLE cells overexpressing MDM4 or those with miR-449b-5p knockdown. Then, tumor weight and tumor volume were measured after mouse sacrifice. Finally, the interaction between miR-449b-5p and MDM4 was explored by Luciferase assay. RESULTS: It was found that MDM4 was upregulated and miR-449b-5p was downregulated in EC tissues. Highly expressed MDM4 and lowly expressed miR-449b-5p were unfavorable to prognosis in EC patients, manifesting as a larger tumor size, more advanced tumor stage and lower overall survival. Besides, overexpression of MDM4 enhanced in vitro proliferative capacity in EC cells and in vivo tumorigenesis in nude mice bearing EC. Similarly, knockdown of miR-449b-5p yielded similar results. Luciferase assay confirmed that MDM4 was the target gene binding to miR-449b-5p, and its level was negatively correlated with miR-449b-5p level in EC. CONCLUSIONS: MiR-449b-5p and MDM4 are downregulated and upregulated in EC species, respectively. They are closely linked to tumor size, tumor stage and overall survival in EC patients. Through negatively regulating MDM4 level, miR-449b-5p inhibits proliferative capacity in EC cells.

[Effect of human stromal vascular fraction gel on the treatment of patients with skin depressed scar and its mechanism].

Objective: To observe content of cytokine in human stromal vascular fraction gel (SVF-GEL) and effect of SVF-GEL on biological behaviors of epidermal and dermal cells in vitro and clinical efficacy of SVF-GEL. Methods: (1) SVF-GEL was prepared using liposuction aspirates harvested from females who received abdomen liposuction in author's unit. SVF-GEL (1 mL) and high-glucose Dulbecco's modified eagle medium (DMEM, 1 mL) were respectively cultured for 24 h with high-glucose DMEM containing 10% fetal calf serum, 10 g/L penicillin, and 10 g/L streptomycin, denoted as SVF-GEL group and negative control group, with 6 samples in each group. Content of epidermal growth factor (EGF) and vascular endothelial growth factor (VEGF) in the supernatant was determined by enzyme-linked immunosorbent assay. (2) A number of 5×10(5) human skin fibroblasts (HSF) and HaCaT cells in logarithmic phase were inoculated and cultured in Transwell chambers for 12 h. All Transwell chambers containing cells were divided into SVF-GEL group (0.5 mL SVF-GEL was added for co-culture) and control group (0.5 mL high-glucose DMEM was added for co-culture), with 9 samples in each group for HSF and HaCaT cells. Scratch assay was performed after culture for 24 h, and residual scratch width was observed at post scratch hour (PSH) 0 (immediately), 24, and 48. Cell migration distance was measured at PSH 24 and 48. After culture for 24, 48, and 72 h, the number of living cell was counted using cell counter. (3) From June 2018 to June 2019, SVF-GEL was applied clinically to treat 15 patients with depressed scars on face, including 2 males and 13 females, aged 19 to 42 years. Survival condition of SVF-GEL and whether complications or not were observed 6 months after surgery. Before surgery and 6 months after surgery, depressed degree, color, and pliability of scar were compared. Vancouver Scar Scale (VSS) was employed to access color, vascularity, and pliability before surgery and 6 months after surgery, and total score was calculated. The number of patients with complete satisfaction or satisfaction was counted six months after surgery. Data were processed with analysis of variance of factorial design, paired samples t test, and Wilcoxon rank sum test. Results: (1) The content of EGF in SVF-GEL group and negative control group was (316.6±12.8) and (3.4±0.6) pg/mL, and the content of VEGF in SVF-GEL group and negative control group was (568.67±12.19) and (4.93±0.16) pg/mL, with statistically significant differences between the two groups (t=48.777, 92.485, P<0.01). (2) Residual scratch widths of HSF and HaCaT in SVF-GEL group and control group were decreased gradually along with time elapse, in which those in SVF-GEL group at PSH 24 and 48 were less than those in control group. At PSH 24 and 48, cell migration distances of HSF and HaCaT in SVF-GEL group were more than those in control group (t(HSF)=-20.304, -43.516, t(HaCaT)=-15.060, -8.684, P<0.01). After culture for 24, 48, and 72 h, the number of living cell of HSF and HaCaT in SVF-GEL group was significantly more than that in control group (t(HSF)=-3.374, -6.809, -18.036, t(HaCaT)=-4.793, -6.028, -8.141, P<0.05 or P<0.01). (3) Six months after surgery, SVF-GEL grafted into patients survived well without complications, and depressed degree of scar ameliorated obviously with lightened pigmentation and softer texture as compared with before surgery. Compared with those before surgery, VSS scores of color, vascularity, and pliability, and total score of 15 patients with depressed scars on face were obviously decreased 6 months after surgery (Z=-2.06, -2.07, -2.07, t=-15.811, P<0.05 or P<0.01). One patient was satisfied with the clinical outcome, and the rest 14 patients were completely satisfied with the clinical outcomes. Conclusions: SVF-GEL contains cytokines EGF and VEGF, which can enhance cell migration ability and proliferation ability of HSF and HaCaT cells and have obvious effects on depressed scars for clinical application.

DIXDC1 prevents oxygen-glucose deprivation/reoxygenation-induced injury in hippocampal neurons in vitro by promoting Wnt/ß-catenin signaling.

OBJECTIVE: Dishevelled-Axin (DIX) domain containing 1 (DIXDC1), a novel DIX domain-containing protein and a positive regulator of Wingless (Wnt) signaling, has previously been reported to play multiple roles in neurodevelopment and neurological disorders. However, whether DIXDC1 plays a role during cerebral ischemia/reperfusion injury remains unknown. In this study, we investigated the potential role of DIXDC1 in neuronal injury induced by oxygen-glucose deprivation and reoxygenation (OGD/R), an in vitro model of cerebral ischemia/reperfusion injury. MATERIALS AND METHODS: Neuronal injury was induced by OGD/R treatment. Relative mRNA expression of DIXDC1 was detected by real-time quantitative polymerase chain reaction (RT-qPCR). Protein expression of DIXDC1 and ß-catenin was determined by Western blot. Cell viability was examined by the cell counting kit-8 assay. Cell cytotoxicity was detected by the lactate dehydrogenase assay. Cell apoptosis was detected by the caspase-3 activity assay. The activity of Wnt/ß-catenin signaling was detected by the luciferase reporter assay. RESULTS: TWe found that DIXDC1 expression was significantly upregulated in hippocampal neurons following OGD/R treatment. Small interfering RNA-mediated silencing of DIXDC1 significantly impaired viability and promoted cell injury and apoptosis in neurons with OGD/R treatment. In contrast, overexpression of DIXDC1 increased the viability and reduced cell injury and apoptosis in neurons with OGD/R treatment, showing protective effects against OGD/R injury. Furthermore, our results showed that DIXDC1 promoted the expression of ß-catenin and activation of Wnt signaling. Notably, inhibition of Wnt signaling significantly abrogated DIXDC-mediated neuroprotective effects. CONCLUSIONS: Our results demonstrate that DIXDC1 prevents OGD/R-induced neuronal injury by promoting Wnt/ß-catenin signaling. Our study indicates that DIXDC1 may play an important role in cerebral ischemia and reperfusion serving as a potential target for the treatment of cerebral ischemia/reperfusion injury.

Effects of hydromorphone and morphine intravenous analgesia on plasma motilin and postoperative nausea and vomiting in patients undergoing total hysterectomy.

OBJECTIVE: To observe the effects of hydromorphone and morphine intravenous analgesia on plasma motilin and postoperative nausea and vomiting in patients undergoing a total hysterectomy. PATIENTS AND METHODS: 80 patients who underwent hysterectomy from April 2015 to June 2016 were randomly divided into two groups, with 40 patients in each group. The two groups received an intravenous infusion of hydromorphone or morphine for analgesia. The VAS pain score and Ramsey sedation score were recorded 4, 8, 12, 24, and 48 hours after the first dose of analgesia. The scores of nausea and vomiting were recorded. The levels of motilin were determined by radioimmunoassay before anesthesia, after anesthesia, during hysterectomy and 1 day after the operation. The results showed that the analgesic effect of hydromorphone was more rapid than morphine. RESULTS: There were significant differences in VAS scores between the two groups at each time point (p<0.05), indicating that the analgesic effect of hydromorphone was better than morphine's one. The scores of Ramsay sedation were less than 6 points at each time point within 48 hours after the operation. The content of plasma motilin in the hydromorphone group was higher than that in the morphine group during the first day after anesthesia. There were 34 cases (85%) of mild nausea and vomiting within 24 hours after the operation in the hydromorphone group. In the morphine group, there were 16 cases (40%) of mild nausea and vomiting within 24 hours after the operation, 10 cases (25%) of severe nausea and vomiting. CONCLUSIONS: The occurrence of severe malignant vomiting after the use of morphine was more than that after the use of hydromorphone. Normal level and function of motilin is the basis of avoiding nausea and vomiting. Too fast or too slow gastrointestinal motility can induce postoperative nausea and vomiting.

Sensitization of mice with wild-type and cold-adapted influenza virus variants: immune response to two H1N1 and H3N2 viruses.

Two A strain influenza viruses, A/Hong Kong/123/77 (A/HK/123/77) (H1N1) and A/Queensland/6/72 (A/Qld/6/72) (H3N2), and the two cold-adapted reassortants which possess the surface antigens of these strains (CR35 and CR6, respectively) were tested for their ability both to induce primary cytotoxic T-cell (Tc cell) responses in mice and to sensitize mice for a second Tc cell response when challenged with a distantly related A strain virus, A/Shearwater/72 (H6N5). After intranasal inoculation, A/Qld/6/72 replicated to higher titers in the lung (1 to 2 log10 50% egg infective doses) than did A/HK/123/77 or either of the reassortants. A/Qld/6/72 induced higher Tc cell responses in the lung than did CR6, and both were more effective than either A/HK/123/77 or CR35 in this respect. When similar doses (10 or 10(3) hemagglutinin units) of each virus were injected intravenously into mice and the spleens were tested for Tc cell activity 6 days later, both A/Qld/6/72 and CR6 were ca. 100-fold better at inducing a primary Tc cell response than A/HK/123/77 or CR35. In contrast, the H1N1 and H3N2 viruses gave rather similar anti-hemagglutinin antibody titers (after intravenous injection) and delayed-type hypersensitivity reactions (after subcutaneous injection). If mice were primed with a low dose of these viruses (10(4) 50% egg infective doses intranasally), A/Qld/6/72 and CR6 were more effective than A/HK/123/77 or CR35 at sensitizing for a secondary Tc cell response when challenged with A/Shearwater/72, but if larger doses were given either intranasally (10(6) 50% egg infective doses) or intravenously (10 to 10(3) hemagglutinin units), all viruses sensitized the mice equally well, despite the fact the A/Shearwater/72 gives a poor primary Tc cell response in mice. Thus, the viral glycoprotein antigens can be important in determining the immunogenicity of the virus and, particularly, the class I antigen-restricted Tc cell response of the host.