Objective To analyze the effects of miR-26a on proliferation,apoptosis,migration and invasion of uveal melanoma cell.Methods The expressions level of miR-26a in the uveal melanoma cell lines SP6.5 and M23 and normal cell line ARPE-19 was detected by qRT-PCR.The SP6.5 cell line was divided into miR-26a mimics group (transfect with miR-26a mimics) and NC group (transfect with scramble by Lipofectamine 2000).The proliferation and apoptosis ability were measured by CCK-8 assay and flow cytometry.The cell wound scratch assay and transwell assay were used to detect the migration and invasion ability.The expression level of enhancer of zeste homolog 2 (EZH2) protein was measured by Western blot.Results The expressions level of miR-26a in uveal melanoma cell line SP6.5 was 0.250 ± 0.029,M23 was 0.350 ±0.017,which was significantly lower than 1.0 in normal cell line ARPE-19 (all P ＜0.001).The OD450 value at 72 hours,96 hours and 120 hours in miR-26a mimics group (0.69 ±--0.09,1.23 ± 0.15,2.12 ± 0.23) were significantly lower than those in NC group (1.39-0.11,2.35 ±0.25,3.53 ±0.27) (all P ＜0.05).The apoptosis rate of miR-26a mimics group (15.60％ ± 2.30％) was significantly higher than that of NC group (5.00％ ± 0.70％) (P ＜ 0.01).The wound healing rate of miR-26a mimics group (23.7％ ±2.1％) was significantly lower than that of NC group (68.9 ±5.1％) (P ＜0.01).While the invasive cell number (45.1 ± 3.9) was also significantly less than NC group (115.3 ± 8.9) (P ＜ 0.01).The expression level of EZH2 protein in miR-26a mimics group (0.39 ±0.09) was significantly lower than that of NC group (1.0,P ＜0.01).Conclusion MiR-26a is low expressed in uveal melanoma cells.Over expression of miR-26a inhibit the proliferation,migration and invasion of uveal melanoma cell,and promote the apoptosis,which may be associated with down-regulated expression of EZH2.

[ ABSTRACT] AIM:To observe the influences of different concentrations of MG132 on apoptosis and beta-amyloid protein ( Aβ) generation in SH-SY5Y cells, and to explore the underlying mechanism.METHODS:SHSY-5Y cells were incubated with MG132 for 24 h.The final concentrations of MG132 were 2.5, 5 and 10μmol/L.The cell viability was de-termined by MTT assay.The cell apoptosis was assessed by flow cytometry.The levels of Aβwere measured by ELISA. The relative protein levels were detected by Western blot.RESULTS:In the SH-SY5Y cells, MG132 reduced the cell via-bility, induced the cell apoptosis, increased the level of Aβ, and increased the expression of the related proteins for Aβgeneration in a concentration-dependent manner.CONCLUSION: MG132 induces apoptosis and increases the levels of Aβ1-42 and Aβ1-40 by regulating the proteins related to Aβgeneration in the SH-SY5Y cells.

Objective:To summarize the clinical and genetic characteristics of children with epilepsy associated with SCN2A gene variations. Methods:A retrospective study was performed. Eleven children with epilepsy admitted to Department of Pediatric Neurology, Linyi People's Hospital from January 2017 to December 2022 were included; all of them had pathogenic SCN2A gene mutation. Genetic results and clinical data as epileptic seizure type/frequency, intelligence and motor development of these 11 children were collected. Epilepsy-related variations and pathogenesis of SCN2A gene were analyzed, and their correlations with clinical phenotypes in these children were analyzed. Results:Among the 11 patients, 6 had self-limited epilepsy (4 with variation in the intracellular domain and 2 in the transmembrane domain), 1 had febrile convulsion accompanied by childhood absent epilepsy (with variation in the intracellular domain), and 4 had developmental epileptic encephalopathy (2 with variation in the extracellular domain and 2 with variation in the transmembrane domain). SCN2A gene was missense mutation in these 11 children, and the mutation site in 6 children was not reported before. Various forms of video EEG discharge were noted, and 1 child with self-limited epilepsy showed transient multifocal epileptic discharge during frequent seizures. Oxcarbazepine and topiramate were effective for self-limiting epilepsy, and lamotrigine was effective in 1 child with late-onset epileptic encephalopathy. Eleven patients were followed up for (66±32) months; the age ranged from 8 months to 11 years and 6 months at the last follow-up; 10 patients had seizure remission and 1 had uncontrolled seizure. Conclusions:Besides self-limited epilepsy and developmental epileptic encephalopathy, SCN2A gene mutations are also associated with febrile convulsion and childhood absent epilepsy. Phenotypic differences are highly correlated with mutation locations; developmental epileptic encephalopathy associated variants are mostly located in extracellular domains, while self-limited epileptic variants are mostly located in intracellular domains.

AIM: To evaluate the bioequivalence of the test and reference formulations of mycophenolate mofetil capsule in Chinese healthy male subjects under fasting and fed conditions. METHODS: This was a 2-treatment, 2-sequence, 4-period, fully replicated crossover study that included 80 Chinese healthy male subjects (40 subjects in the fasting group and 40 subjects in the fed group, respectively). Subjects were assigned to receive a single oral administration of the test or reference formulation at a dose of 0.25 g in each period. The plasma concentration of mycophenolate mofetil (MMF) and metabolite mycophenolic acid (MPA) were analyhed by LC-MS/MS. The major pharmacokinetic parameters of MMF and MPA were calculated using non-compartmental analysis by WinNonlin 8.0. The statistical analysis was performed by SAS 9.4. Average bioequivalence (ABE) analysis was applied where it has been demonstrated that the within-subject standard deviation of the reference formulation (S

The clinical utilization of doxorubicin (Dox) in various malignancies is restrained by its major adverse effect: irreversible cardiomyopathy. Extensive studies have been done to explore the prevention of Dox cardiomyopathy. Currently, ferroptosis has been shown to participate in the incidence and development of Dox cardiomyopathy. Sorting Nexin 3 (SNX3), the retromer-associated cargo binding protein with important physiological functions, was identified as a potent therapeutic target for cardiac hypertrophy in our previous study. However, few study has shown whether SNX3 plays a critical role in Dox-induced cardiomyopathy. In this study, a decreased level of SNX3 in Dox-induced cardiomyopathy was observed. Cardiac-specific Snx3 knockout (Snx3-cKO) significantly alleviated cardiomyopathy by downregulating Dox-induced ferroptosis significantly. SNX3 was further demonstrated to exacerbate Dox-induced cardiomyopathy via induction of ferroptosis in vivo and in vitro, and cardiac-specific Snx3 transgenic (Snx3-cTg) mice were more susceptible to Dox-induced ferroptosis and cardiomyopathy. Mechanistically, SNX3 facilitated the recycling of transferrin 1 receptor (TFRC) via direct interaction, disrupting iron homeostasis, increasing the accumulation of iron, triggering ferroptosis, and eventually exacerbating Dox-induced cardiomyopathy. Overall, these findings established a direct SNX3-TFRC-ferroptosis positive regulatory axis in Dox-induced cardiomyopathy and suggested that targeting SNX3 provided a new effective therapeutic strategy for Dox-induced cardiomyopathy through TFRC-dependent ferroptosis.