Zhou J.J.,Guiyang Medical University
Nan fang yi ke da xue xue bao = Journal of Southern Medical University | Year: 2011
To study the effects of neuron specific enolase (NSE) gene silencing on the cell proliferation and apoptosis of lung cancer cells in vitro. NSE protein expression was detected in human small cell lung cancer cell line NCI-H446 and non-small cell lung cancer cell line A549 by immunocytochemistry, and a small interference RNA (siRNA) was transfected into the cells to inhibit NSE gene expression. The changes in the cell cycle, apoptosis, Ki67 protein and caspase-3 activity in the transfected cells were observed by flow cytometry, Western blotting and colorimetric assay, respectively. Both A549 and NCI-H446 cells expressed NSE protein. Transfection of siRNA for NSE gene significantly inhibited the expression of NSE gene in the cells, resulting in an inhibition rate exceeding 90%. NSE gene silencing caused significantly decreased cell percentage in S phase and the expression of Ki67 protein, and increased the cell apoptotic rate and caspase-3 activity. NSE gene expression promotes the cell proliferation and inhibits the cell apoptosis in lung cancer cells with neuroendocrine differentiation, which might be a causative factor contributing to increased malignancy of the cells.
Yan Z.,Columbia University |
Yan H.,Zhejiang University |
Ou H.,Guiyang Medical University
Gene | Year: 2012
The liver performs a vital role in metabolic process, which makes it an attractive target organ for gene therapy. To improve the effects of gene therapy in disorders caused by metabolic disturbance, we quantitatively evaluated six promoters, CMV, EF1α, PGK, apoE, thyroxine binding globulin (TBG), and cytochrome P450 2E1 (CYP2E1) by measuring the expression of α1-antitrypsin, which is controlled by these promoters and introduced via a lentivirus-mediated delivery system in the liver. The results showed that the TBG promoter presents as highly active though in general it is slightly lower than the ubiquitous CMV and EF1α. The expression of exogenous genes driven by the TBG promoter demonstrates to be much higher than by PGK, apoE, and CYP2E1 promoters, and the fragment of -. 435. bp to -. 26. bp from transcription start site (TSS) in the TBG promoter region is identified as the optimum region to direct transgene expression at a higher level. In addition, we further confirmed that the TBG promoter confers transgene persistent and specific expression within the liver up to several months after integration. The data suggests that the TBG promoter is a valuable tool and will greatly facilitate the optimization of vector design in hepatic gene therapy. © 2012 Elsevier B.V.
Luo W.,Guiyang Medical University
Zhongguo xiu fu chong jian wai ke za zhi = Zhongguo xiufu chongjian waike zazhi = Chinese journal of reparative and reconstructive surgery | Year: 2012
To construct a new type of self-assembling peptide nanofiber scaffolds-RGDmx, and to study the cell compatibility of the new scaffolds and the proliferation and chondrogenic differentiation of precartilaginous stem cells (PSCs) in scaffolds. PSCs were separated and purified from newborn Sprague Dawley rats by magnetic activated cell sorting and indentified by immunohistochemistry and immunofluorescent staining. The RGDmx were constructed by mixing KLD-12 and KLD-12-PRG at volume ratio of 1:1. PSCs at passage 3 were seeded into the KLD-12 scaffold (control group) and RGDmx scaffold (experimental group). The proliferation of PSCs in 2 groups were observed with the method of cell counting kit (CCK)-8 after 1, 3, 7, and 14 days after culture. The RGDmx were constructed by mixing KLD-12-PRG and KLD-12 at different volume ratios of 0, 20%, 40%, 60%, 80%, and 100% and the proliferation of PSCs was also observed. The complete chondrogenic medium (CCM) was used to induce chondrogenic differentiation of PSCs in different scaffolds. The differentiation of PSCs was observed by toluidine blue staining and RT-PCR assay. PSCs were separated and purified successfully, which were identified by immunohistochemistry and immunofluorescent staining methods. The results of CCK-8 showed that the absorbance (A) value in the experimental group increased gradually and reached the highest at 7 days; the A value in the experimental group was significantly higher than that in the control group at 7 days and 14 days (P < 0.05). Meanwhile, the A value in the RGDmx scaffold with a volume ratio of 40% was significantly higher than those in others (P < 0.05). After 14 days of induction culture with CCM, the toluidine blue staining results were positive in 2 groups; the results of RT-PCR showed that the expression levels of collagen type II and the aggrecan in the experimental group were significantly higher than those in the control group (P < 0.05). The self-assembling peptide nanofiber scaffold-RGDmx is an ideal scaffold for tissue engineer because it has good cell compatibility and more effective properties of promoting the differentiation of PSCs to chondrocytes.
Chen J.,Guiyang Medical University
Ceramics International | Year: 2015
As one of the most potential negative electrode materials, Na2Ti6O13 is expected to play an important role in the area of high-performance battery. In this work, we have developed an easy, efficient and controllable method to prepare rod-shaped Na2Ti6O13 crystals. This approach utilized a single-source molten salt strategy and only needed to sinter a special precursor synthesized from an aqueous solution containing H3BO3 and (NH4)2TiF6 in presence of sodium salts. The component and shape of precursor crystals can be tuned by adjusting the reagent concentration and reaction temperature. By sintering precursor crystals in air at 900 °C for 30 min, Na2Ti6O13 with high crystallinity and purity can be obtained. X-ray diffraction and scanning electron micrographs results of different sintering times show that the sintering process can be divided into two steps. Firstly, the precursor crystals are converted to TiO2 (anatase) nano-particles and amorphous sodium salts. Subsequently, molten salt reaction occurs between amorphous sodium salts and TiO2 and forms rod-shaped Na2Ti6O13 crystals. © 2015 Elsevier Ltd and Techna Group S.r.l. All rights reserved.
Wang Q.,Guiyang Medical University
Zhonghua wei chang wai ke za zhi = Chinese journal of gastrointestinal surgery | Year: 2012
To compare surgical efficacy after three different reconstruction techniques after radical resection of distal gastric cancer. Clinical data of 169 cases of distal gastric cancer operated in our hospital from 2007 to 2010 were retrospectively analyzed. The reconstruction techniques included Billroth I (anastomosis (n=60), Billroth II (anastomosis (n=41), and Roux-en-Y anastomosis (n=68). Efficacy among 3 groups was compared. Specific symptoms scale was used to evaluate the quality of life in three methods after three months. Compared to Billroth I(anastomosis and Billroth II (anastomosis, Roux-en-Y anastomosis had longer operative time [(266.3±70.4) min vs. (196.2±54.3) min, and (228.5±67.7) min], more blood loss [(220.9±67.6) ml vs. (170.5±61.5) ml and (188.5±76.7) ml], and shorter time to gastric tube removal [(2.6±1.5) d vs. (3.1±1.3) d and (3.6±1.2) d], milder postoperative reflux and heartburn sensation(specific symptoms scale, 1.8±0.4 vs. 1.9±0.6 and 2.6±0.4, P<0.05). Although Roux-en-Y anastomosis is not consistent with physiological route and the procedure is more complex to perform, it can effectively prevent reflux complications. Roux-en-Y anastomosis is a better reconstruction technique after radical resection of distal gastric cancer.