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猪细胞因子IL-2、IL-4和IFN-γ作为免疫增强剂和新型基因工程疫苗的研究 被引量:1
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作者 王洪光 汤德元 +5 位作者 李春燕 曾智勇 罗险峰 甘振磊 刘建 郝飞 《猪业科学》 2014年第1期99-101,共3页
通过综述猪IL-2、IL-4和IFN-γ的基因结构、作为免疫佐剂和作为构建新型基因工程疫苗方面研究进展,以了解细胞因子IL-2、IL-4和IFN-γ的相关研究成果,为细胞因子进行更深入的研究提供参考。
关键词 猪细胞因子 IL-2 IL-4 IFN-Γ 免疫增强剂 新型基因工程疫苗
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猪GM-CSF基因的克隆及其真核表达载体的构建 被引量:1
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作者 黄海军 高其双 +9 位作者 钱运国 邵淑敏 黄远全 陈志华 陶弼菲 向敏 占才耀 夏瑜 王连芳 任远 《生物技术通报》 CAS CSCD 北大核心 2011年第4期112-115,132,共5页
采用PCR方法从猪外周血液淋巴细胞cDNA中扩增出与预期设计大小相符的GM-CSF基因特异性条带,PCR产物经EcoR I和Xho I双酶切后,插入到载体pIRES2-EGFP构建成真核表达载体pIRES2-EGFP-GM-CSF。经PCR鉴定、限制性内切酶酶切分析和克隆片段... 采用PCR方法从猪外周血液淋巴细胞cDNA中扩增出与预期设计大小相符的GM-CSF基因特异性条带,PCR产物经EcoR I和Xho I双酶切后,插入到载体pIRES2-EGFP构建成真核表达载体pIRES2-EGFP-GM-CSF。经PCR鉴定、限制性内切酶酶切分析和克隆片段序列测定、比较,证实了重组质粒的正确性。将构建好的真核表达质粒转染到山羊胎儿成纤维细胞中进行瞬时表达,荧光检测证实细胞转染成功。pIRES2-EGFP-pGM-CSF真核表达载体的成功构建为下一步在细胞水平研究GM-CSF蛋白功能以及进一步将其开发为高档疫苗佐剂奠定了基础。 展开更多
关键词 细胞-巨噬细胞集落刺激因子 克隆 真核表达载体 瞬时转染
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稳定转染pGM-CSF基因的哺乳动物细胞系的建立
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作者 陶弼菲 高其双 +8 位作者 陈志华 向敏 占才耀 钱运国 夏瑜 华娟 王莲芳 任远 黄海军 《中国兽医杂志》 CAS 北大核心 2013年第8期6-8,12,共4页
构建稳定表达猪pGM-CSF基因的细胞系可以为工业化生产猪pGM-CSF并将其应用于新型兽药的开发提供生产细胞源。本试验用脂质体介导法将真核表达载体plRES2-EGFP-pGM-CSF瞬时转染BHK-21细胞,通过不同浓度的G418加压筛选和进一步采用单个克... 构建稳定表达猪pGM-CSF基因的细胞系可以为工业化生产猪pGM-CSF并将其应用于新型兽药的开发提供生产细胞源。本试验用脂质体介导法将真核表达载体plRES2-EGFP-pGM-CSF瞬时转染BHK-21细胞,通过不同浓度的G418加压筛选和进一步采用单个克隆法筛选,建立稳定转染且高效表达pGM-CSF的BHK-21细胞株9个,采用Elisa方法测定稳定转染pGM-CSF的BHK-21细胞株细胞培养上清液中pGM-CSF的表达水平达到329.37±13.76ng/mL。为下一步从细胞培养物中大量提取目的蛋白并最终开发出一种用于提高动物疫苗免疫效力的新型生物制剂奠定基础。 展开更多
关键词 细胞-巨噬细胞集落刺激因子 哺乳动物细胞 稳定转染 蛋白表达
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Inhalation of hydrogen gas reduces liver injury during major hepatotectomy in swine 被引量:5
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作者 Lei Xiang Jing-Wang Tan +5 位作者 Li-Jie Huang Lin Jia Ya-Qian Liu Yu-Qiong Zhao Kai Wang Jia-Hong Dong 《World Journal of Gastroenterology》 SCIE CAS CSCD 2012年第37期5197-5204,共8页
AIM: To study the effect of H2 gas on liver injury in massive hepatectomy using the Intermittent Pringle maneuver in swine. METHODS: Male Bama pigs (n = 14) treated with ketamine hydrochloride and Sumianxin Ⅱ as ... AIM: To study the effect of H2 gas on liver injury in massive hepatectomy using the Intermittent Pringle maneuver in swine. METHODS: Male Bama pigs (n = 14) treated with ketamine hydrochloride and Sumianxin Ⅱ as induc- tion drugs followed by inhalation anesthesia with 2% isoflurane, underwent 70% hepatotectomy with loss of bleeding less than 50 mL, and with hepatic pedicle occlusion for 20 min, were divided into two groups: Hydrogen-group (n = 7), the pigs with inhalation of 2% hydrogen by the tracheal intubation during major hepa- totectomy; Contrast-group (n= 7), underwent 70% hepatotectomy without inhalation of hydrogen. Hemo- dynamic changes and plasma concentrations of alanine aminotransferase (ALT), aspartate aminotransferase (AST), hyaluronic acid (HA), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and malondialdehyde (MDA) in liver tissue were measured at pre-operation, post-hepatotectomy (PH) 1 h and 3 h. The apoptosis and proliferating cell nuclear antigen (PCNA) expres- sion in liver remnant were evaluated at PH 3 h. Then we compared the two groups by these marks to evalu- ate the effect of the hydrogen in the liver injury during major hepatotectomy with the Pringle Maneuver in the swine. RESULTS: There were no significant differences in body weight, blood loss and removal liver weight be- tween the two groups. There was no significant differ- ence in changes of portal vein pressure between two groups at pre-operation, PH 30 min, but in hydrogen gas treated-group it slightly decrease and lower than its in Contrast-group at PH 3 h, although there were no significant difference (P = 0.655). ALT and AST in Hydrogen-group was significantly lower comparing to Contrast-group (P = 0.036, P = 0.011, vs P = 0.032, P = 0.013) at PH 1 h and 3 h, although the two groups all increased. The MDA level increased between the two group at PH i h and 3 h. In the hydrogen gas treated- group, the MDA level was not significantly significant at pre-operation and significantly low at PH 1 h and 3 h comparing to Contrast-group (P = 0.0005, P = 0.0004). In Hydrogen-group, the HA level was also significantly low to Contrast-group (P = 0.0005, P = 0.0005) al- though the two groups all increased at PH 1 h and 3 h. The expression of cluster of differentiation molecule 31 molecules Hydrogen-group was low to Contrast-group. However, PCNA index (%) was not statistically signifi- cant between the two groups (P = 0.802). Micropho- tometric evaluation of apoptotic index (AI) in terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling-stained tissue after hepatotectomy for 3h, the AI% level in the hydrogen was significantly low to Contrast-group (P = 0.012). There were no significant difference between Hydrogen-group and Contrast- group at pre-operation (P = 0.653, P = 0.423), but after massive hepatotectomy, the TNF-α and IL-6 levels increase, and its in Hydrogen-group was significantly low compared with Contrast-group (P = 0.022, P = 0.013, vs P = 0.016, P= 0.012), respectively. Hydro- gen-gas inhalation reduce levels of these markers and relieved morphological liver injury and apoptosis.CONCLUSION: H2 gas attenuates markedly ischemia and portal hyperperfusion injury in pigs with massive hepatotectomy, possibly by the reduction of inflamma- tion and oxidative stress, maybe a potential agent for treatment in clinic. 展开更多
关键词 Massive hepatotectomy Hydrogen gas An-ti-oxidant HYPERPERFUSION MALONDIALDEHYDE OXIDATIVESTRESS
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