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Objective:To screen and identify differentially expressed long non-coding RNA (lncRNAs) in peripheral blood of children with sepsis, and to explore the role of lncRNAs in the pathogenesis of sepsis in children.Methods:The peripheral blood samples of 3 children with sepsis admitted to the ICU of Children′s Hospital of Capital Institute of Pediatrics from January to December 2016 and 3 healthy children who underwent physical examination in our hospital during the same period were selected, and the differential expression profiles of lncRNAs and mRNAs were screened by lncRNAs sequencing technology.The target genes of differentially expressed lncRNAs were predicted and the relationship pairs of lncRNA-mRNA related to F 1F O-ATPase activity were constructed according to the results of GO analysis.Further increasing the sample size, we verified the expression of some F 1F O-ATPase activity-related mRNAs and lncRNAs which were differentially expressed in the screening results by real-time fluorescent quantitative polymerase chain reaction(qRT-PCR). Results:Sequencing results showed that there were 252 lncRNAs with significant differential expression in peripheral blood of children with sepsis compared with healthy children, of which 86 were up-regulated and 166 were down-regulated; meanwhile, there were 2 652 mRNAs with significant differential expression, of which 955 were up-regulated and 1 697 were down-regulated.The results of qRT-PCR showed that the expression of lncRNA ENST00000621933.1, ENST00000616950.1 and ENST00000595748.1 in peripheral blood of children with sepsis increased( P<0.05), while the expression of MT-ATP8, ATP5E and ENST00000624705.1, ENST00000615535.1 in peripheral blood of children with sepsis decreased( P<0.05), which was consistent with the sequencing results. Conclusion:lncRNAs are differentially expressed in peripheral blood of children with sepsis compared with healthy children.The expression levels of lncRNA ENST00000621933.1, ENST00000616950.1, ENST00000595748.1, ENST00000624705.1 and ENST00000615535.1 which their target genes are MT-ATP8 and ATP5E may be related to the development of sepsis in children.
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AIM:To observe the effects of the combination of berberin and epirubicin on the cell cycle of T24 bladder cancer cells and the underlying mechanisms.METHODS:The cancer cells were exposed to epirubicin in the presence or absence of different concentrations of berberin.The viability of the cancer cells was determined by MTT assay.The cell cycle distribution was detected by flow cytometry, and the protein levels of cyclin D1, CDK2, CDK4, P21 and P27 were detected by Western blot.RESULTS:Berberine markedly enhanced the inhibitory effect of epirubicin on the viability of T24 cells and promoted epirubicin-induced cell cycle arrest at G0/G1 phase as compared with the negative control cells.Epirubicin increased the protein expression of P27 and P21, both of which were enhanced by treatment with berberin.In contrast, berberin exposure further decreased the protein expression of cyclin D1, CDK2 and CDK4 in epirubicin-treated T24 cells.CONCLUSION:Berberine significantly promotes epirubicin-induced G0 /G1 phase arrest in human bladder cancer cells by up-regulating P27 and P21 expression and inhibiting the expression of cyclin D1, CDK2 and CDK4.
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AIM: To observe the effects of berberine (Ber) on enterocyte apoptosis in septic mice and its pos-sible mechanism.METHODS: Male C57BL/6 mice (8 ~10 weeks old) were randomly divided into sham group, cecal ligation and puncture (CLP) group, CLP +Ber group and sham +Ber group.The mice in CLP group underwent CLP ope-ration, and the mice in sham groups suffered a similar operation except the ligation and puncture.After the sham or CLP operation, the mice were administered intragastrically with distilled water or berberine (50 mg/kg) within 2 h.After 20 h, the mice were killed with excess pentobarbital sodium and the ileum tissues were removed.The histological changes of the intestine were observed and the enterocyte apoptosis was examined by determining the protein level of cleaved caspase-3. Furthermore, mitochondrial Bax, cytoplasm cytochrome C (Cyt C) and the total proteins of Bcl-2, Fas, FasL and Fas-as-sociated protein with death domain (FADD) were examined by Western blot.The mRNA expression of tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DBH) was measured by real-time PCR.RESULTS: The extensive ileum injuries, including remarkably increased leukocytes and necrosis of intestinal villus were observed 20 h after CLP.In CLP group, the protein levels of cleaved caspase-3, cytoplasm Cyt C, as well as Fas, FasL were significantly increased, but the Bcl-2 level was decreased.Bax translocation into mitochondria was promoted.However, FADD was not changed significantly.The mRNA expression of TH and DBH was also increased sharply in CLP group.On the contrary, treatment with berberine made a considerable alleviating alteration in the ileum of the septic mice.CONCLUSION: Treatment with berberine pro-vides protective effects on intestinal injury in septic mice by reducing enterocyte apoptosis, and its possible mechanism may be involved in the inhibition of the endogenous and exogenous apoptosis pathways.
ABSTRACT
Sepsis is a complex pathophysiological disorder arising from a systemic inflammatory response to infection.It is considered as a major cause of death among critically ill patients in intensive care units.Mitochondrial dysfunction and impaired oxidative phosphorylation have been implicated in the pathogenesis of sepsis.The mitochondrial respiratory chain complex Ⅴ is the key enzyme of oxidative phosphorylation.It is response for ATP synthesis and there is a reduction of complex Ⅴ activity in the early sepsis.However,in the late sepsis,under ischemic conditions, it accounts for significant ATP hydrolysis and the activity is up-regulation.Here,this review focuses on the changes of complex Ⅴ activity,mechanism of regulation and therapy in sepsis.