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Objective: The pathogenesis of diabetes is closely related to islet β-cell dysfunction. Lnc-NTF3-5 participates in the occurrence and development of various diseases. However, Lnc-NTF3-54's effect on islet β-cell dysfunction in high glucose environment remains unclear., Materials and Methods: The islet β cell MIN6 cells were cultured in vitro and randomly divided into control group, high glucose group, NTF3-5 siRNA group, and NTF3-5 group, which was respectively transfected with Lnc-NTF3-5 siRNA and Lnc-NTF3-5 plasmid under high glucose condition. Lnc-NTF3-5 expression was measured by real time PCR and cell proliferation was assessed by MTT assay. In addition, Caspase 3 activity, SOD activity, and ROS content were also detected along with the secretion of IL-10 and IL-1 by enzyme-linked immunosorbent assay (ELISA)., Results: Compared with control group, Lnc-NTF3-5 expression in MIN6 cells was significantly increased in high glucose environment (p<0.05). In high glucose environment, Lnc-NTF3-5 plasmid transfection up-regulated Lnc-NTF3-5 expression, inhibited cell proliferation, increased Caspase 3 activity, and decreased SOD activity. Meanwhile, Lnc-NTF3-5 plasmid also increased ROS content and IL-1 level, and decreased IL-10 level and insulin secretion. Compared with high glucose group, the differences were statistically significant (p<0.05). However, transfection of Lnc-NTF3-5 siRNA down-regulated Lnc-NTF3-5 expression under high glucose environment and reversed the above changes. Compared with high glucose group, the differences were statistically significant (p<0.05)., Conclusions: Lnc-NTF3-5 expression is increased in high glucose environment. Targeting Lnc-NTF3-5 can inhibit islet cell apoptosis, oxidative stress, and promote islet cell proliferation and insulin secretion.

Objective: The aim of this study was to investigate the effect of micro-ribonucleic acid (miR)-130a on neuronal injury in rats with intracerebral hemorrhage (ICH) through the phosphatase and tensin homolog deleted on chromosome ten/phosphatidylinositol 3-hydroxy kinase/protein kinase B (PTEN/PI3K/AKT) signaling pathway., Materials and Methods: A total of 30 healthy male rats were randomly divided into three groups, including the blank control group, ICH model group (ICH group) and ICH model + miR-130a treatment group (miR-130a treatment group). The differences in neurological injury, the number of apoptotic cells in brain tissues, the activity of Caspase-9 and protein expressions of PTEN/PI3K/AKT were analyzed among the three groups, respectively., Results: Neurological function was normal without injury in the control group. However, the neurological injury was severe in the ICH group and mild in the miR-130a treatment group. There were statistically significant differences in neurological function in the control group relative to those of the ICH group and miR-130a treatment group (p<0.05). Meanwhile, the neurological injury was markedly milder in the miR-130a treatment group than that of the ICH group, showing a statistically significant difference (p<0.05). The number of apoptotic cells was remarkably smaller in the control group when compared with the ICH group and miR-130a treatment group. However, it was markedly larger in the ICH group than that of the miR-130a treatment group, showing significant differences (p<0.05). The activity of Caspase-9 was significantly lower in the control group than ICH group and miR-130a treatment group (p<0.05). However, it increased remarkably in the ICH group compared with that of the miR-130a treatment group (p<0.05). Moreover, the protein level of PTEN in the ICH group was significantly higher than control group and miR-130a treatment group, displaying statistically significant differences (p<0.05). However, no marked difference in the protein level of PTEN was observed between the control group and miR-130a treatment group (p>0.05). The protein levels of the phosphorylated 3-hydroxy kinase (p-PI3K) and phosphorylated protein kinase B (p-AKT) were remarkably lower in the ICH group than those of the control group and miR-130a treatment group, displaying statistically significant differences (p<0.05). However, they were remarkably higher in the miR-130a treatment group than that of the control group (p<0.05)., Conclusions: MiR-130a promotes neuronal growth in brain tissues in ICH rats and alleviates neuronal injury after ICH through the PTEN/PI3K/AKT signaling pathway. Our findings suggest that miR-130a exerts important clinical significance in the treatment of ICH.

Objective: To investigate the effects of neutrophils alkaline phosphatase (NAP) on the migration, reactive oxygen species (ROS) generation and apoptosis of neutrophil-like differentiated HL-60 cells., Methods: NAP was overexpressed in HL-60 cells via transfecting coding sequence of NAP by lentivirus. The effectivity of NAP overexpression was confirmed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. HL-60 cells were differentiated into neutrophil-like cells by exposure to 1.5% DMSO. The migration and ROS generation of neutrophil-like cells with NAP overexpression were detected by transwell migration test and flow cytometry, respectively. Cell apoptosis were detected by flow cytometry. The expression of apoptosis-related protein Bax, caspase-3 and caspase-9 in neutrophil-like cells were observed by Western blotting after NAP overexpression., Results: Over 80% HL-60 cells presented green fluorescence after GFP-NAP infection by lentivirus and screening by puromycin. In addition, the levels of both gene and protein of NAP were up-regulated in these cells. After 5-day culture with 1.5% DMSO, the bulk of induced cell was smaller and the surface appeared many bumps and irregular shape. The ratio of nuclear and cytoplasmic decreased and nucleolus disappeared. The nuclear chromatin changed from dense to loose and the nuclear morphology appeared to be rod and segmented. The percentage of CD11b(+) cell increased from 26.25% to 98.55%. The transwell migration test showed that the number of migrated cells was higher in neutrophil-like cells with NAP overexpression compared with the negative control [(15.30±3.65) ×10(3) vs (8.00±0.78) ×10(3)] (P<0.001). RESULTS of flow cytometry suggested that the mean fluorescence intensity (MFI) of intracellular ROS was significantly higher in neutrophil-like cells with NAP overexpression compared with the negative control (355.70 ± 20.10 vs 103.22 ± 4.71) (P< 0.001). In addition, Western blotting showed that the expressions of apoptosis-related protein Bax, active-caspase-3 and active-caspase-9 were all up-regulated in neutrophil-like cells with NAP overexpression compared with the negative control., Conclusion: NAP could promote the migration and ROS generation of neutrophil-like cells and accelerate the cell apoptosis.

Expression of apoptotic protease activating factor-1 (Apaf-1) gradually decreases during brain development, and this decrease is likely responsible for the decreased sensitivity of brain tissue to apoptosis. However, the mechanism by which Apaf-1 expression is decreased remains elusive. In the present study, we found that four microRNAs (miR-23a/b and miR-27a/b) of miR-23a-27a-24 and miR-23b-27b-24 clusters play key roles in modulating the expression of Apaf-1. First, we found that miR-23a/b and miR-27a/b suppressed the expression of Apaf-1 in vitro. Interestingly, the expression of the miR-23-27-24 clusters in the mouse cortex gradually increased in a manner that was inversely correlated with the pattern of Apaf-1 expression. Second, hypoxic injuries during fetal distress caused reduced expression of the miR-23b and miR-27b that was inversely correlated with an elevation of Apaf-1 expression during neuronal apoptosis. Third, we made neuronal-specific transgenic mice and found that overexpressing the miR-23b and miR-27b in mouse neurons inhibited the neuronal apoptosis induced by intrauterine hypoxia. In conclusion, our results demonstrate, in central neural system, that miR-23a/b and miR-27a/b are endogenous inhibitory factors of Apaf-1 expression and regulate the sensitivity of neurons to apoptosis. Our findings may also have implications for the potential target role of microRNAs in the treatment of neuronal apoptosis-related diseases.

Compound C, a well-known inhibitor of AMP-activated protein kinase (AMPK), has been reported to induce apoptosis in some types of cells. However, the underlying mechanisms remain largely unclear. Using a DNA microarray analysis, we found that the expression of many genes was downregulated upon treatment with compound C. Importantly, compound C caused transcriptional repression with the induction of p53, a well-known marker of transcriptional stress response, in several cancer cell lines. Compound C did not induce the phosphorylation of p53 but dramatically increased the protein level of p53 similar to some other transcriptional inhibitors, including 5,6-dichloro-1-β-D-ribobenzimidazole (DRB). Consistent with previous reports, we found that compound C initiated apoptotic death of cancer cells in an AMPK-independent manner. Similar to DRB and actinomycin D (ActD), two classic transcription inhibitors, compound C not only resulted in the loss of Bcl-2 and Bcl-xl protein but also induced the phosphorylation of eukaryotic initiation factor-alpha (eIF2α) on Ser51. Hence, the phosphorylation of eIF2α might be a novel marker of transcriptional inhibition. It is noteworthy that compound C-mediated apoptosis of cancer cells is correlated with decreased expression of Bcl-2 and Bcl-xl and the phosphorylation of eIF2α on Ser51. Remarkably, compound C exhibits potent anticancer activities in vivo. Taken together, our data suggest that compound C may be an attractive candidate for anticancer drug development.

A new method based on photoelectric measurement for analyzing apoptosis of cell-free MCF-7 nucleoli is reported. Supported bilayer lipid membrane (s-BLM) was used to enclose nucleoli in biological environment. The s-BLM was self-assembled on the wall of a super-thin cell. During the apoptosis induced by Taxol, the photoelectric current of the self-assembled s-BLM/nucleoli was found decreasing with time, suggesting the degradation of nucleus DNA. Electron transfer along the DNA double helix and along nuclear skeleton is assumed in the interpretation. This novel photoelectric analytical method may provide a rapid and sensitive technique to evaluate apoptosis.

Micro RNAs (mi RNAs) are one of a growing class of noncoding RNAs that are involved in the regulation of a wide range of metabolic processes including cellular differentiation, cell proliferation and apoptosis. The generation of mi RNA is regulated in complex ways, for example by small interfering RNAs (small nucleolar and nuclear RNAs) and various other metabolites. This complexity of control is likely to explain how a relatively small part of the DNA that codes for proteins has enabled the evolution of such complex organisms as mammals. Non-protein-coding DNA is therefore thought to carry the memory of early evolutionary steps that led to progressively complex metabolic controls. Clinically, mi RNAs are becoming increasingly important following the recognition that some congenital abnormalities can be traced to defects in mi RNA processing. The potential for manipulating metabolism and affecting disease processes by the pharmaceutical or biological targeting of specific mi RNA pathways is now being tested. mi RNAs are also released into the extracellular milieu after packaging by cells into nano-sized extracellular vesicles. Such vesicles can be taken up by adjacent and possibly more distant cells, thereby allowing coordinated intercellular communication in specific tissues. Extracellular mi RNAs found in the blood stream may also serve as novel biomarkers for both diagnosing specific forms of cancer and assessing the likelihood of metastasis, and as powerful prognostic indices for various cancers. Here, we discuss the role of intracellular and extracellular mi RNAs in nutritional control of various (patho)physiological processes. In this review, we provide an update of the presentations from the 25th Marabou Symposium (Stockholm, 14-16 June 2013) entitled 'Role of mi RNA in health and nutrition', attended by 50 international experts. FOR A COMPILATION OF THE GENERAL DISCUSSION AT THE MARABOU SYMPOSIUM, click here [ABSTRACT FROM AUTHOR]