No abstract

Effect of vitamin D on apoptosis of peripheral blood T-lymphocyte subsets in treatment of neonatal sepsis was investigated. A total of 150 neonatal patients with sepsis were randomly divided into vitamin D treatment group (observation group) and treatment control group, while 100 healthy newborns were selected as healthy control group. T-lymphocyte subsets were detected by flow cytometer, the levels of tumor necrosis factor-α, interleukin-1 and calcitonin were determined by double-antibody immunoluminometric assay, and the effect of vitamin D on the above indicators in the treatment of sepsis was observed. Serum 25(OH)D (22.52±5.56 mg/l) in the treatment group was obviously increased compared with that in the treatment group (14.85±6.14 mg/l) (P<0.05), but the levels in the two groups were remarkably lower than that in the normal control group (26.38±6.56 mg/l), and the differences were statistically significant (P<0.05). Cluster of differentiation 4 (CD4+) T-lymphocyte subset in sepsis patients was obviously reduced compared with that in the healthy control group (P<0.01); the difference in comparison of CD8+ T-lymphocyte subset between sepsis patients and healthy people was not statistically significant (P>0.05). After treatment for 72 h, CD4+ T-lymphocytes were increased, and the ratio of CD4+ to CD8+ was close to 1, suggesting that the effect was superior to that in the treatment control group. The inflammatory factor levels in children with sepsis were evidently higher than those in the healthy control group (P<0.01), and high-level states of inflammatory factors were significantly improved after treatment with vitamin D for 72 h, indicating that the effect was superior to that in the treatment group. The results indicated that the prognosis of sepsis patients treated with vitamin D is improved, and the mechanism may be achieved by regulating T-lymphocyte subsets and inflammatory factors.

Background: Toll-like receptor4 (TLR4) has proven to be an important factor that's responsible for the development of postoperation infection. MicroRNAs (miRNAs) are widely regarded as key mediators of gene expression. The objectives of our study were to identify miRNA(s) and the target genes differentially expressed in monocytes in the individuals with postoperation infection.Methods: MiRNA microarrays were performed to identify and compare miRNA expression in monocytes from those with or without postoperative infection. In-silico analysis was used to further investigate the target miRNAs and finally, luciferase assay and real-time polymerase chain reaction (PCR) were performed to confirm the target miRNA identified. Enzyme-linked immunosorbent assay, real-time PCR and Western-blot were performed to explore the role of miR-140 involved in postoperation infection. Results: MiRNA microarray results showed that ten miRNAs were upregulated in the postoperation infection group, while six miRNAs were downregulated, compared with those in the postoperation group without infection. Computational analysis was further performed to reveal that four miRNAs (miR-140, miR-7, miR-448, and miR-217) targeted the 3′-untranslated region (UTR) of TLR4 mRNA. The luciferase assay showed that only miR-140 inhibited luciferase activity of wild-type TLR4 3′-UTR and the luciferase activity of the cells cotransfected with miR-7, miR-448 or miR-217 and wild-type or mutant TLR4 3′-UTR was comparable with the control. Furthermore, only miR-140 levels were significantly lower in the postoperation infection group, while levels of miR-217, miR-7, and miR-448 showed no obvious difference between the postoperation infection and postoperation without infection groups. TLR4, tumor necrosis factor-α (TNF-α), and IL-6 levels were much higher in the postoperation infection group. In comparison with the control group, TLR4, TNF-α and Interleukin 6 (IL-6) levels in cells were decreased following transfection with miR-140 mimics and TLR4 small interfering RNA. However, the cells treated with lipopolysaccharides increased TLR4, TNF-α, and IL-6 levels.

The hormonally active form of vitamin d (Vd), 1,25-dihydroxyvitamin d3, has been reported to be a key immunoregulator in the reduction of inflammation. In this study, we investigated the effects of Vd in an experimental sepsis cell model, and the underlying mechanisms. The sepsis cell model was first established in monocytes, isolated from newborns and healthy adults, which were stimulation with lipopolysaccharide (LPS). We observed that cell viability was significantly impaired in the monocytes after LPS stimulation, using a cell counting Kit-8 and trypan blue assays. Additionally, ELISA revealed that LPS stimulation significantly elevated the expression of interleukin 6 (IL-6), IL-10 and tumor necrosis factor-α (TNF-α). The expression levels of Toll-like receptor (TLR4), myeloid differentiation primary response gene 88 (Myd88), and Toll-IL-1 resistance-domain-containing adapter-inducing interferon-β (TRIF) mRNA were also significantly elevated under LPS stimulation using reverse transcription-quantitative PcR and western blot analysis. VD treatment could significantly suppress the effects of LPS simulation on monocytes by negatively regulating inflammatory cytokines and TLR4/Myd88/TRIF signaling. Furthermore, a regulatory feedback mechanism was proposed to involve TLR4, Myd88 and TRIF in the sepsis cell model. In conclusion, VD may effectively decrease the release of inflammatory cytokines by inhibiting the TLR4/Myd88/TRIF signaling pathway, could be considered as a potential therapeutic agent for the treatment of sepsis.

Background This study aimed to explore the regulatory relationship between growth arrest special 5 (GAS5) and interleukin‐1β (IL‐1β) implicated in the development of febrile seizure (FS). Method The presence of FS and the genotype of GAS5 were used as two different indicators to divide the 50 newborn babies, recruited in this study, into different groups. The potential regulatory relationship among GAS5, miR‐21, and IL‐1β was identified by measuring their expression using quantitative reverse‐transcription polymerase chain reaction and immunohistochemistry assays among different sample groups. Computational analyses and luciferase assays were also conducted to verify the interaction between GAS5, miR‐21, and IL‐1β. Result GAS5 and IL‐1β expression was upregulated in cells collected from FS patients or genotyped as INS/DEL and DEL/DEL, whereas the expression of miR‐21 was decreased in above samples, indicating a negative relationship between miR‐21 and GAS5/IL‐1β. Results of the computational analysis showed that miR‐21 directly bound to and increased the expression of GAS5, whereas the expression of IL‐1β was suppressed by miR‐21. In the presence of GAS5, the expression of miR‐21 was lowered, whereas the expression of IL‐1β was increased. Conclusion The results obtained in this study supported the conclusion that GAS5 negatively regulated the expression of miR‐21, which in turn negatively regulated the expression IL‐1β. Therefore, the overexpression of GAS5 could decrease the magnitude of FS.