Establishment of L-periaxin gene knock-out RSC96 cell line / 生物工程学报

Periaxin, a protein of noncompact myelin, is specifically expressed in the peripheral nervous system (PNS). There are two protein isoform L-periaxin and S-Periaxin by alternative splicing of periaxin gene, playing an important role in the initiation of myelin formation. So far, 18 different mutation sites in L-periaxin gene have been found to induce the peripheral demyelinating neurological charcot-marie-tooth diseases subtype 4F (CMT4F). The technique of activation of transcription activator-like effector nucleases (TALENS) was used to knock out the L-periaxin gene in RSC 96 cell line of Rattus. According to the design principle, the knock-out site of L-periaxin was assured to NLS domain of L-periaxin, which is target sequence of left and right arms of TALEN. The knock-out vectors of TALEN-L and TALEN-R were established and transfected into RSC96 cell. After puromycin screening, L-periaxin was knocked out successfully in RSC96 cell, which is confirmed by DNA sequence. The mutation efficiency is 21.6%. S-periaxin, not L-periaxin can be detected by Western blotting in L-periaxin gene knock-out RSC96 cell. The cell growth rate was decreased and the number of cells in G1 increased and decreased in S phase in L-periaxin gene knock-out RSC96 cell by flow cytometry and MTT assay.

Multivariate analysis of prognostic makers in patients with nasopharyngeal carcinoma intreatment of radiotherapy combined with induction chemotherapy / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To observe the therapeutic effect of radiotherapy combined with TPF in patients with nasopharyngeal carcinoma (NPC) and to explore the influencing factors on prognosis of NPC.@*METHOD@#A retrospective analysis of 144 patients with nasopharyngeal carcinoma receiving radiotherapy combined with chemotherapy, from January 2006 to December 2011, was conducted. The survival analysis in patients with nasopharyngeal carcinoma were performed. The impact of age, sex, clinical stage, alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), lactate dehydrogenase (LDH), homocysteine (Hcy), uric acid, NK cell proportion, T cell proportion, Carbohydrate antigen on survival time was observed with univariate and multivariate methods.@*RESULT@#By the last follow-up, 9 cases were lost . The follow-up rate was 93.75 percent and the average follow-up time were 54.67 ±17.17 (37-88) months. The age, clinical stage, LDH, proportion of T cells and NK cells were independent factor in patients with NPC with univariate analysis (P < 0.01). COX regression multivariate analysis revealed that age, clinical stage and LDH were strong risk factor affecting prognosis (P < 0.01) and the proportion of NK cells and T cells were protective factors (P < 0.01).@*CONCLUSION@#Radiotherapy combined with TPF is more effective than radiotherapy alone in patients with nasopharyngeal carcinoma . Age, clinical stage and LDH may adversely influence the disease prognosis, and NK cells and T cell proportion may act as protective factors.