Objective To investigate the role and mechanism of the zinc finger protein Kruppel-like transcription factor 9 (KLF9) in the stimulation of type I interferon expression induced by herpes simplex virus type 1 (HSV-1) in macrophages. Methods Agarose Gel electrophoresis, quantitative real-time PCR (qRT-PCR) and western blot analyses were employed to detect the KLF9 relative expression in bone marrow-derived macrophages (BMDMs) from Klf9^-/- (gKO) mice and wild-type (WT) mice. RNA-seq analysis was utilized to identify the potential targeted genes upon HSV-1 stimulation in BMDMs. ELISA was used to measure the potent of IFN-β in the supernatant of BMDMs derived from gKO and WT mice after HSV-1 stimulation. qRT-PCR analysis was employed to further confirm the changes of Ifnb1 and interferon-stimulated gene (ISG) such as interferon-induced protein with tetratricopeptide repeats 1 (Ifit1), interferon-stimulated exonuclease gene 20 (Isg20), cholesterol 25-hydroxylase (Ch25h) and 2'-5' oligoadenylate synthetase-like 1 (Oasl1). Western blot was used to detect the expression of phosphorylated interferon regulatory factor-3 (p-IRF3), IRF3, phosphorylated interferon regulatory factor-7 (p-IRF7), IRF7, phosphorylated nuclear factor-kappa B p65 (p-NF-κB p65) and NF-κB p65. CUT-Tag and ChIP-qPCR assay were utilized to confirm the binding region of KLF9 in Ifnb1. Results The KLF9 expression was significantly decreased in BMDMs from gKO mice compared with that from WT mice. The RNA-seq analysis showed that Klf9 deletion in BMDMs resulted in an impaired type I interferon signaling pathway. The qRT-PCR analysis revealed that Klf9 deletion in BMDMs led to a significant decrease of Ifnb1 and ISG such as Ifit1, Ch25h and Oasl1 except Isg20. Moreover, ELISA revealed that Klf9 knockout in BMDMs resulted in a significant decrease of IFN-β secreted from BMDMs. Mechanistically, KLF9 directly binds to the promoter of Ifnb1. Conclusion KLF9 is essential for macrophages to resist HSV-1 infection.
Animals ; Kruppel-Like Transcription Factors/physiology* ; Interferon-beta/metabolism* ; Macrophages/virology* ; Mice ; Herpesvirus 1, Human/physiology* ; Mice, Knockout ; Signal Transduction ; Mice, Inbred C57BL ; Interferon Regulatory Factor-3/genetics* ; Interferon Regulatory Factor-7/genetics* ; Gene Expression Regulation

Objective To explore rationale and clinical application of simplified modified radical thyroideetomy for differentiated thyroid carcinoma.Methods From Jan.2007 to Jun.2010,349 cases of differentiated thyroid carcinoma received simplified operative procedure based on standard modified radical thyroidectomy.The simplified procedure took a low small collar incision(about 10-12 cm).In separating upper and lower skin flaps,subcutaneous tissues covering posterior triangle of neck and posterior edge of sternoeleidomastoid muscle were spared to protect sensory nerves.Subtotal thyroidectomy Was performed to resect the affected lobe,isthmus,and the majority of opposite lobe without considering the size of primary tumor or whether metastasis to the neck lymph nodes happened.Soft tissues of the mainly metastatic areas(Ⅱ a、Ⅲ、Ⅳ、Ⅴb)were cleared.The accessory nerve was not exposed routinely to avoid stimulation.Lymph nodes metastasis in different areas was recorded respectively.Complications in different operative modes were compared.Results Compared with standard modified radical thyroidectomy,the simplified mode had shorter scar-and no limit of neck mobility.Because of muscles and nerves pemervation,movement dysfunction and abnormal sensation of neck and shoulder decreased obviously.The operation duration was shortened.Cervical lymph node status Was evaluated,which provided basis for prognosis judgment and comprehensive treatment.Conclusions The simplified modified radical procedure has the benefit of decreased trauma while maintains the similar recurrence rate compared to modified radical thyroidectomy.It improvs the life quality of patients.This procedure fits the principle of functional radical neck dissection better.

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