Development and validation of a prognostic nomogram for patients with ganglioneuroblastoma: A SEER-based study.

Background: The objective of this study was to construct a prognostic nomogram for ganglioneuroblastoma (GNB), as the prognosis of GNB is difficult to accurately predict before therapy. Methods: The data were collected from the Surveillance, Epidemiology, and End Results (SEER) database. The patients included in this study were randomly divided into a development group and a validation group at a ratio of 7:3. Univariate and multivariate Cox regression analyses were used to filter the variables. Receiver operating characteristic (ROC) curves and calibration curves were used to assess the nomogram. All patients were redivided into two groups based on their nomogram total points, and overall survival was compared. Results: A total of 1194 GNB patients were retrospectively included, with 835 and 359 patients in the development and validation groups, respectively. Five independent prognostic factors, including age, primary tumor site, SEER stage, surgery and chemotherapy, were screened out and included in the nomogram. The consistency index (C-index) of the Cox regression model was 0.862 and 0.827 in the development group and the validation group, respectively. The areas under the receiver operating characteristic (ROC) curve (AUC) showed that the nomogram had good accuracy in predicting 3-, 5- and 10-year overall survival for GNB patients. The calibration curves of the nomogram showed good agreement between the predicted outcomes and the actual observations. The Kaplan-Meier (KM) survival curves revealed that patients with nomogram scores below the median had a better prognosis. Conclusions: Age, primary tumor site, SEER stage, surgery and chemotherapy may be independent prognostic factors for GNB. We constructed a nomogram based on the SEER database to predict the prognosis of GNB, but further optimization by adding more risk factors is needed for clinical application.

Association between age at first birth and postpartum depression: A two-sample mendelian randomization analysis.

Background: Previous observational research has documented an association between age at first childbirth (AFB) and postpartum depression (PPD). However, the causal relationship remains unclear. This study aimed to assess the causal effects of AFB on PPD using a two-sample Mendelian randomization (MR) analysis. Methods: Three sets of instrumental variables were obtained from the United Kingdom Biobank (UK Biobank), Neale Lab consortium and a meta-analysis of genome-wide association studies (GWAS). Single-nucleotide polymorphisms (SNPs) associated with the PPD phenotype were obtained from the Finngen consortium, which included 13,657 cases and 236,178 controls. Inverse variance weighted (IVW), weighted median, weighted mode, and MR-Egger methods to evaluate causal effects. Heterogeneity was assessed using Cochran's Q test and funnel plots. Horizontal pleiotropy and sensitivity were assessed using the MR-Egger intercept test and "leave-one-out" analysis, respectively. Further meta-analysis was performed to validate the robustness of this relationship. Additionally, the potential mediating effects of risk factors associated with PPD were analyzed. Results: Strong causal effects between AFB and PPD was found in both IVW and weighted median methods, which was further supported by meta-analysis (IVW, odds ratio [OR] 0.59 [95% confidence interval (CI) 0.36-0.96, p = 0.03]; weighted median, OR 0.59 [95% CI 0.37-0.95, p = 0.03]). The power of the MR supports the robustness of the findings. Heterogeneity or horizontal pleiotropy was not observed. Major depressive disorders, family income levels, and marital stress were identified as potential mediating factors in the causal relationships. Conclusion: Results of MR analysis supported the causal effect of increased AFB in reducing the risk for PPD.

Hyperbranched Cationic Glycogen Derivative-Mediated IκBα Gene Silencing Regulates the Uveoscleral Outflow Pathway in Rats.

The role of the IκB/NF-κB signaling pathway in the uveoscleral outflow pathway was investigated with IκBα gene silencing mediated by the 3-(dimethylamino)-1-propylamine-conjugated glycogen (DMAPA-Glyp) derivative. The IκBα-siRNA-loaded DMAPA-Glyp complex was transfected into the ciliary muscles of rats by intracameral injection (labeled as the DMAPA-Glyp+siRNA group). The Lipofectamine™ 2000 (Lipo)/siRNA complex and the naked siRNA were set as the controls. The mRNA and protein expression of IκBα, NF-κBp65, and MMP-2 were analyzed by real-time PCR, western blotting, and in situ gelatin zymography. Nuclear translocation of NF-κBp65 was analyzed by immunofluorescence. Rat intraocular pressure (IOP) was monitored pre- and postinjection. Gene transfection efficiency and toxicity of the DMAPA-Glyp derivative were also evaluated. After RNA interference (RNAi), IκBα mRNA and protein expression were significantly inhibited. NF-κBp65 mRNA and protein expression showed no significant differences. Nevertheless, nuclear translocation of NF-κBp65 occurred in the DMAPA-Glyp+siRNA group. Both mRNA expression and activity of MMP-2 increased, with the largest increase in the DMAPA-Glyp+siRNA group. IOP in the DMAPA-Glyp+siRNA group fell to the lowest level on day 3 after RNAi. The levels of Cy3-siRNA in the ciliary muscle of the DMAPA-Glyp+siRNA group did not significantly decrease over time. At 7 and 14 d after RNAi, no significant pathological damage was detectable in the eyes injected with the DMAPA-Glyp derivative or the DMAPA-Glyp/siRNA complex. Taken together, our results suggest that downregulation of IκBα expression in the ciliary muscle plays a crucial role in reducing the IOP values of rats. IκBα may become a new molecular target for lowering IOP in glaucoma. The DMAPA-Glyp derivative is safe and feasible as an effective siRNA vector in rat eyes.

In Vitro Screening and Transfection Concentration Optimization of Cynomolgus Monkey IκBα-siRNA.

PURPOSE: To seek for a small interfering RNA (siRNA) sequence targeting a cynomolgus monkey inhibitor of nuclear factor kappa B α (IκBα) that can specifically and effectively suppress IκBα gene expression of cynomolgus monkey ciliary muscle (CM) cells and trabecular meshwork (TM) cells in vitro and screen for optimal siRNA transfection concentration. METHODS: Three IκBα-specific double-stranded siRNAs were designed and synthesized. They were transfected into primarily cultured cynomolgus monkey CM cells and TM cells. The mRNA and protein levels of IκBα were examined by using real-time quantitative polymerase chain reaction (real-time PCR) and western blot to screen a pair of candidate valid sequences with the highest inhibitory rate. Both cells were transfected with Cy5-labeled nonspecific control-siRNA (NC-siRNA) of four different concentrations (10, 20, 50, and 100 nmol/L(nM)), and flow cytometry was used to assess transfection efficiency. Then, cells were transfected with the candidate valid IκBα -siRNA of the same four concentrations, and the cytotoxicity was detected by using Cell Counting Kit-8 (CCK8), and the inhibitory efficiency of IκBα was identified via real-time PCR to find out optimal siRNA transfection concentration. RESULTS: The suppression effect of the siRNA targeting the GCACTTAGCCTCTATCCAT of IκBα gene was most obvious by in vitro screening. The inhibitory rate of IκBα was 82% for CM cells and 82% for TM cells on the mRNA level and 98% for CM cells and 93% for TM cells on the protein level, respectively. The results of flow cytometry showed that the transfection efficiency was the highest at 100 nM, which was 89.0% for CM cells and 48.2% for TM cells, respectively. The results of CCK8 showed that there was no statistically significant difference in cell viability after transfection of different concentrations of IκBα-siRNA. The results of real-time PCR indicated that there was no statistical difference in the inhibitory efficiency of IκBα after transfection of different concentrations of IκBα-siRNA. CONCLUSION: It proves that the siRNA targeting the GCACTTAGCCTCTATCCAT of IκBα gene is the valid sequence to suppress cynomolgus monkey IκBα expression of CM cells and TM cells by RNAi. 10 nM is the optimal transfection concentration.