Some transcription factors that specifically bind double-stranded DNA appear to also function as RNA-binding proteins. Here, we demonstrate that the transcription factor Sox2 is able to directly bind RNA in vitro as well as in mouse and human cells. Sox2 targets RNA via a 60-amino-acid RNA binding motif (RBM) positioned C-terminally of the DNA binding high mobility group (HMG) box. Sox2 can associate with RNA and DNA simultaneously to form ternary RNA/Sox2/DNA complexes. Deletion of the RBM does not affect selection of target genes but mitigates binding to pluripotency related transcripts, switches exon usage and impairs the reprogramming of somatic cells to a pluripotent state. Our findings designate Sox2 as a multi-functional factor that associates with RNA whilst binding to cognate DNA sequences, suggesting that it may co-transcriptionally regulate RNA metabolism during somatic cell reprogramming.
By combining asymmetric PCR and overlap extension, we developed a novel asymmetric overlap extension PCR (AOE-PCR) method for site-directed mutagenesis which bypassed the need for intermediate purification and excluded the amplification of a wild-type template. This method was used to introduce single base mutations into a small GTPase gene from cotton and to simultaneously introduce two mutations just by repeating this method using the first round AOE-PCR products as template. Our results suggested that the AOE-PCR method represents a valuable improvement of the original overlap extension PCR for site-directed mutagenesis.
The present study was conducted to evaluate the effect of thymectomy during open heart surgery on immunological function of T lymphocytes in the treatment of children with congenital heart disease (CHD). No significant difference was found in the sjTREC level between pre-thymectomy and post- thymectomy in the non-thymectomy group and the small partial resection group (P>0.05) However, the sjTREC level decreased from the pre-surgical level at 1 month (P<0.01) and 12 months (P<0.01) in the sub-total resection group. No differences were found in proportions of CD3, CD4 and CD8 lymphocytes, proliferative ability of lymphocytes and expression of IL-2, IL-4 and IFN-γ after surgery between controls and three groups of patients (P>0.05). In the sub-total resection group, respiratory infection frequency (4.7±1.7 times) did not differ significantly from control group one year after surgery (P>0.05); however, mean days of anti-infection were significantly increased (P less than 0.01). In conclusion, sub-total thymectomy leads to a decrease in the sjTREC level in CHD children, whereas the function of peripheral mature T lymphocytes is normal.
Objective: To explore the clinical characteristics of and analyze the risk factors for hydrocephalus in children with bacterial meningitis. Methods: Retrospective study of a sample of children with bacterial meningitis seen on the pediatric service of Shengjing Hospital of China Medical University between January 1, 2010, and December 31, 2016. Results: Overall, 9.36% (25/267) of patients presented with hydrocephalus. Among patients with hydrocephalus, the age at onset of bacterial meningitis was usually <6 months, 15 patients had confirmed bacterial etiology, and 1 patient died. The most significant results of multivariate analysis for hydrocephalus were a rural living situation, altered level of consciousness, previous treatment with antibiotics, initial cerebrospinal fluid protein >2 g/L, C-reactive protein >100 mg/L, and dexamethasone use. Conclusions: A severe clinical manifestation and significant laboratory index at admission are the most important predictors of hydrocephalus in children with bacterial meningitis.
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