AIM The purpose of this study was to evaluate the diagnostic value of trefoil factor family 3(TFF3) for the early detection of colorectal cancer(CC). METHODS Serum TFF3 and carcino-embryonic antigen(CEA) were detected...AIM The purpose of this study was to evaluate the diagnostic value of trefoil factor family 3(TFF3) for the early detection of colorectal cancer(CC). METHODS Serum TFF3 and carcino-embryonic antigen(CEA) were detected in 527 individuals, including 115 healthy control(HC), 198 colorectal adenoma(CA), and 214 CC individuals in the training group. RESULTS Serum TFF3 showed no significant correlation with age, gender, or tumor location but showed significant correlation with the tumor stage. Serum TFF3 in the CC group was significantly higher than in the HC or CA group. The AUC values of TFF3 for discriminating between HC and CC and between CA and CC were 0.930(0.903, 0.958) and 0.834(0.796, 0.873). A multivariate model combining TFF3 and CEA was built. Compared to TFF3 or CEA alone, the multivariate model showed significant improvement(P < 0.001). For discriminating between HC and CC, HC and early stage CC, HC and advanced stage CC, CA and CC, CA and early stage CC, and CA and advanced stage CC in the training group, the sensitivities were 92.99%, 91.46%, 93.18%, 73.83%, 76.83%, and 81.82%, and the specificities were 91.30%, 91.30%, 93.91%, 88.38%, 77.27%, and 88.38%, respectively. After validation, the sensitivities were 89.39%, 85.71%, 90.79%, 72.73%, 71.43%, and 78.95%, and the specificities were 87.85%, 87.85%, 2.52%, 87.85%, 80.77%, and 87.50%, respectively. CONCLUSION The multivariate diagnostic model that included TFF3 and CEA showed significant improvement over the conventional biomarker CEA and might provide a potential method for the early detection of CC.展开更多
目的构建含有人三叶因子3(trefoil factor family 3,TFF3)基因的腺病毒载体,并观察其对胆管上皮细胞迁移的调控作用。方法利用PCR技术对目的基因TFF3全长进行扩增,测序后亚克隆至腺病毒穿梭质粒构建载体pAdTrack-CMV-TFF3,转化至含腺病...目的构建含有人三叶因子3(trefoil factor family 3,TFF3)基因的腺病毒载体,并观察其对胆管上皮细胞迁移的调控作用。方法利用PCR技术对目的基因TFF3全长进行扩增,测序后亚克隆至腺病毒穿梭质粒构建载体pAdTrack-CMV-TFF3,转化至含腺病毒骨架质粒pAd-Easy的大肠杆菌株BJ5183感受态细胞中进行同源重组,筛选重组子并经内切酶PacⅠ线性化,转染293细胞进行病毒包装及扩增,并进行滴度测定。构建的TFF3腺病毒载体感染人胆管上皮细胞后用RT-PCR及Western blot检测目的基因表达水平,并进行划痕实验观察细胞迁移能力的变化。结果成功扩增了大小约为458 bp的人TFF3基因,测序报告显示目的基因序列与NCBI报道一致;成功构建了TFF3的同源重组腺病毒穿梭载体,经293细胞包装及扩增的病毒滴度为1.3×109CPU/mL。RT-PCR及Western blot结果显示转染人胆管上皮细胞后其TFF3表达明显增强,细胞迁移能力也明显高于对照组。结论成功构建含TFF3基因的腺病毒载体,为进一步研究TFF3在胆管上皮损伤修复中的作用奠定了基础。展开更多
BACKGROUND： Sepsis has become the greatest threat to in-patients, with a mortality of over 25%.The dysfunction of gut barrier, especially the immunological barrier, plays an important role in the development of sepsi...BACKGROUND： Sepsis has become the greatest threat to in-patients, with a mortality of over 25%.The dysfunction of gut barrier, especially the immunological barrier, plays an important role in the development of sepsis. This dysfunction occurs after surgery, but the magnitude of change does not differentiate patients with sepsis from those without sepsis. Increased intestinal permeability before surgery is of no value in predicating sepsis. The present study aimed to observe the changes of intestinal mucosal immunologic barrier in rat models of sepsis induced by cecal ligation and puncture.METHODS： Sixty Sprague-Dawley rats were randomly divided into a sepsis group （n=45） and a control group （n=15）. The rats in the sepsis group were subjected to cecal ligation and puncture （CLP）, whereas the rats in the control group underwent a sham operation. The ileac mucosa and segments were harvested 3, 6 and 12 hours after CLP, and blood samples were collected. Pathological changes, protein levels of defensin-5 （RD-5） and trefoil factor-3 （TFF3） mRNA, and lymphocytes apoptosis in the intestinal mucosa were determined. In an additional experiment, the gut-origin bacterial DNA in blood was detected.RESULTS： The intestinal mucosa showed marked injury with loss of ileal villi, desquamation of epithelium, detachment of lamina propria, hemorrhage and ulceration in the sepsis group. The expression of TFF3 mRNA and level of RD-5 protein were decreased and the apoptosis of mucosal lymphocyte increased （P〈0.05） in the sepsis group compared with the control group. Significant differences were observed in RD-5 and TFF3 mRNA 3 hours after CLP and they were progressively increased 6 and 12 hours after CLP in the sepsis group compared with the control group （P〈0.05, RD-5 F=11.76, TFF3 F=16.86 and apoptosis F=122.52）. In addition, the gut-origin bacterial DNA detected in plasma was positive in the sepsis group.CONCLUSION： The immunological function of the intestinal mucosa was impaired in septic rats and further deteriorated in the course of sepsis.展开更多
基金Supported by The Capital Health Development Special Scientific Research Projects,No.2014-2-2154National Natural Science Foundation of China,No.81471761 and No.81501568
文摘AIM The purpose of this study was to evaluate the diagnostic value of trefoil factor family 3(TFF3) for the early detection of colorectal cancer(CC). METHODS Serum TFF3 and carcino-embryonic antigen(CEA) were detected in 527 individuals, including 115 healthy control(HC), 198 colorectal adenoma(CA), and 214 CC individuals in the training group. RESULTS Serum TFF3 showed no significant correlation with age, gender, or tumor location but showed significant correlation with the tumor stage. Serum TFF3 in the CC group was significantly higher than in the HC or CA group. The AUC values of TFF3 for discriminating between HC and CC and between CA and CC were 0.930(0.903, 0.958) and 0.834(0.796, 0.873). A multivariate model combining TFF3 and CEA was built. Compared to TFF3 or CEA alone, the multivariate model showed significant improvement(P < 0.001). For discriminating between HC and CC, HC and early stage CC, HC and advanced stage CC, CA and CC, CA and early stage CC, and CA and advanced stage CC in the training group, the sensitivities were 92.99%, 91.46%, 93.18%, 73.83%, 76.83%, and 81.82%, and the specificities were 91.30%, 91.30%, 93.91%, 88.38%, 77.27%, and 88.38%, respectively. After validation, the sensitivities were 89.39%, 85.71%, 90.79%, 72.73%, 71.43%, and 78.95%, and the specificities were 87.85%, 87.85%, 2.52%, 87.85%, 80.77%, and 87.50%, respectively. CONCLUSION The multivariate diagnostic model that included TFF3 and CEA showed significant improvement over the conventional biomarker CEA and might provide a potential method for the early detection of CC.
基金This study was supported by the National Natural Science Foundation of Guangdong Province （06021323）.
文摘BACKGROUND： Sepsis has become the greatest threat to in-patients, with a mortality of over 25%.The dysfunction of gut barrier, especially the immunological barrier, plays an important role in the development of sepsis. This dysfunction occurs after surgery, but the magnitude of change does not differentiate patients with sepsis from those without sepsis. Increased intestinal permeability before surgery is of no value in predicating sepsis. The present study aimed to observe the changes of intestinal mucosal immunologic barrier in rat models of sepsis induced by cecal ligation and puncture.METHODS： Sixty Sprague-Dawley rats were randomly divided into a sepsis group （n=45） and a control group （n=15）. The rats in the sepsis group were subjected to cecal ligation and puncture （CLP）, whereas the rats in the control group underwent a sham operation. The ileac mucosa and segments were harvested 3, 6 and 12 hours after CLP, and blood samples were collected. Pathological changes, protein levels of defensin-5 （RD-5） and trefoil factor-3 （TFF3） mRNA, and lymphocytes apoptosis in the intestinal mucosa were determined. In an additional experiment, the gut-origin bacterial DNA in blood was detected.RESULTS： The intestinal mucosa showed marked injury with loss of ileal villi, desquamation of epithelium, detachment of lamina propria, hemorrhage and ulceration in the sepsis group. The expression of TFF3 mRNA and level of RD-5 protein were decreased and the apoptosis of mucosal lymphocyte increased （P〈0.05） in the sepsis group compared with the control group. Significant differences were observed in RD-5 and TFF3 mRNA 3 hours after CLP and they were progressively increased 6 and 12 hours after CLP in the sepsis group compared with the control group （P〈0.05, RD-5 F=11.76, TFF3 F=16.86 and apoptosis F=122.52）. In addition, the gut-origin bacterial DNA detected in plasma was positive in the sepsis group.CONCLUSION： The immunological function of the intestinal mucosa was impaired in septic rats and further deteriorated in the course of sepsis.
