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91.
92.
目的:构建同时携带低氧诱导因子-1α(HIF-1α)和角质细胞生长因子(KGF)N腺病毒载体(pAdxsi-GFP-HIF-KGF),观察其在防治肺损伤潜在的应用前景。方法:低氧处理A549细胞后提取总RNA并逆转录为eDNA作为模板,依据GeneBank公布的HIF-1α cDNA设计引物,并分别引入KpnI和BamHI酶切位点,PCR扩增后将目的基因HIF-1α连接到载体pShuttle-CMV-EGFP上,构建重组质粒pShuttle-GFP—HIF。然后以质粒plRES2-EGFP-KGF为模板,用引入NheI和PmeI酶切位点的引物PCR扩增KGF基因并克隆到重组质粒pShuttle-GFP-HIF上,获得穿梭质粒重组质粒pShuttle—GFP-HIF—KGF。采用细菌内重组方法将目的序列重组到pAdxsi病毒骨架栽体上构建携带HIF.10t和KGF双基因的重组腺病毒载体pAdxsi-GFP-HIF-KGF。检测重组腺病毒滴度后,转染人肺泡上皮细胞A549,检测目的基因的转染表达。结果:通过对构建质粒克隆进行测序及酶切,证实携带HIF—lot和KGF双基因的重组腺病毒载体pAdxsi-GFP-HIF-KGF构建成功,且构建的重组腺病毒纯度好、滴度高。用pAdxsi-GFP-HIF-KGF以100MOI转染A549细胞后24h后在荧光显微镜下可观察到细胞有较强的绿色荧光表达,48h时荧光更强;转染48hELISA法检测培养上清中HIF-1蛋白表达水平为(56.36±4.53)ng/mL,KGF蛋白表达水平为(60.20±2.92)ng/mL。结论:成功构建了腺病毒栽体pAdxsi-GFP-HIF-KGF,其转染效率及目的基因的蛋白表达水平较高,具有潜在的进一步在肺损伤局部应用的前景,为后期制备可以同时发挥KGF、HIF-1作用的基因治疗药物打下基础,同时为高海拔地区应激性急性肺损伤的有效防治提供实验基础。 相似文献
93.
基因枪在大鼠闭塞性血管病基因治疗中的应用 总被引:1,自引:0,他引:1
探讨利用基因枪技术转移肝细胞生长因子基因治疗大鼠肢体闭塞性血管病的可行性,构建了携带人肝细胞生长因子(HGF)基因的重组真核表达载体(pUDKH),在制备好大鼠下肢闭塞性血管病模型后,通过基因枪或肌肉直接注射法,向局部缺血部位肌肉中转移pUDKH,每只5ug(基因枪)和12ug(肌肉注射),应用常规组织病理切片(H.E.染色)及免疫组织化学方法观察血管形成及基因表达,转移pUDKH后第10天,用基因枪和直接注射法转移的局部肌肉组织的HGF的表达明显高于转移空白质粒(pUDK)的对照组,pUDKH组可见明显的小血管新生,而pUKD组至20天时仍未观察到或仅见到极少量的新生血管,基因枪与肌肉注射两组相比血管密度无明显差异,采用基因枪直接转移pUDKH裸露质粒子肢体缺血局部的方法是可行的,转移的基因可在局部有效表达,达到促进血管形成的预期目的。 相似文献
94.
京津冀风沙源区沙化土地治理关键技术研究与示范 总被引:2,自引:0,他引:2
京津冀风沙源区是我国北方生态屏障的重要组成部分。面向我国北方风沙区沙化土地综合治理、典型脆弱生态修复与保护等重大科技需求,京津冀风沙源区沙化土地治理关键技术研究与示范项目将综合运用长期定位观测、控制实验、多源遥感数据融合、技术示范等方法,重点研究京津冀风沙源区土地沙化形成机制与生态修复机理,研发一批沙化土地治理与产业化关键技术,并在各治理区开展试验示范,集成京津冀风沙源区沙化土地治理与产业化技术体系,构建沙化土地综合整治空间决策支持系统,为京津风沙源治理工程建设、保障京津冀地区生态安全及满足2022年北京冬奥会生态需求提供科技支撑。 相似文献
95.
In order to show the function of agricultural protection of biodiversity at the level of homegarden, comparative analysis was operated between 20 farmer households through random sampling and farmers recommended in Horqin Left Wing Real Banner Tongliao city Inner Mongolia Autonomous Region and 10 farmer households of the Han nationality in the neighborhood. Thirty sampling plots of homegardens are investigated. The result indicates there are 100 homegarden plants in Bashou village, 84 in Hariwusu livestock farm, and 87 in Gongjihao village. It involves the cultivated and wild vegetables, fruits, and plants used for ornamental, medicine and forage. The comparative analysis of species richness demonstrates, among these villages the species richness of Bashou village is highest, while Hariwusu livestock farm is lowest. The former village contributes greatly to the conservation of plant biodiversity. These differences result from their respectively different natural conditions, homegarden areas, homegarden history, homegarden management conditions, traditional knowledge and culture understanding of homegarden manager, local farmers and herdsmen on homegarden plants.The homegarden plants from the three investigated villages are cataloged preliminary in the end of the paper. 相似文献
96.
Amiodarone (AM) is an effective anti-arrhythmic drug. We investigated the role of mast cells and macrophages on AM induced pulmonary fibrosis and the action of atorvastatin on this fibrosis. Rats were allocated into four groups; negative control (1), positive control (2), 30 mg/kg body weight/day AM (3) and AM + 10 mg/kg/day atorvastatin (4). Lungs were harvested and prepared for histology and immunohistochemistry. Hematoxylin and eosin stained sections of group 3 exhibited disorganized lung architecture. We found cellular debris in the lumen of both intrapulmonary bronchi and bronchioles with partial disruption of the thickened epithelial lining and mononuclear cellular infiltration into the lamina propria. We also observed thickening of the epithelial lining and the smooth muscle layer. Congested, dilated and thickened blood capillaries and thickened inter-alveolar septa were observed with mononuclear cellular infiltrates in the lung of group 3. Most alveoli were collapsed, but some dilated ones were detected. In some alveoli, type ?? pneumocytes were increased, while type I cells were decreased. We observed significant increases in the amount of collagen in the thickened inter-alveolar septa, around bronchioles and around blood capillaries in sections from group 3. We found a significant increase in mast cells and alveolar macrophages in group 3 compared to group 1. Mast cells and macrophages appear to play important roles in AM induced pulmonary fibrosis. Atorvastatin appears to attenuate this condition. 相似文献
97.
The local herdsmen were served as informants, methods of interviews and voucher specimen collection and identification have been used to conduct ethnobotanical field investigations on wild edible plants in the Xilingol typical steppe area, Inner Mongolia. The results show that 29 species and two varieties of wild plants used for food and drinks by the Mongolians in Xilingol typical steppe area. The edible parts of the plants are whole plant, aerial parts, roots, stems, bulbs, leaves, flowers, fruits or/and seeds respectively. Among them, the leaf is the most widely used part. Six categories of food uses based on the mode of folk edible use were established to classify wild food plants, including wild grain, vegetables, fruit, substitute for tea, seasoning, and snacks. Among them, vegetables were the largest group, followed by plants used as substitute for tea, and plants used for seasoning. Wild plants are usually eaten raw or cooked by the local people. Raw materials and prepared food from wild plants are preserved through the method of pickling and drying. A distinguishing feature of region and ethnic group is that mutton, beef, fresh milk and yoghourt are necessarily used in cooking dish and making milk tea from wild plants by local Mongolians. 相似文献
98.
本文运用量子力学方法对蛋白质分子中孤波传播的非线性动力学特征进行了探讨。研究表明:电离辐射产生的自由基对蛋白质分子的伤害将会对携带能量、信息的孤立子波传播产生较为显著的影响。 相似文献
99.
Janny C de Grauw Chris HA van de Lest Paul René van Weeren 《Arthritis research & therapy》2009,11(2):R35-8
Introduction
Inflammation is an important feature of many joint diseases, and levels of cartilage biomarkers measured in synovial fluid may be influenced by local inflammatory status. Little is known about the magnitude and time course of inflammation-induced changes in cartilage tissue turnover as measured in vivo by synovial fluid markers. We aimed to study temporal changes in concentrations of inflammatory mediators, matrix metalloproteinase activity and cartilage biomarkers over 1 week in joints with experimentally induced inflammation. 相似文献100.
CC van Diemen DS Postma M Siedlinski A Blokstra HA Smit HM Boezen 《Respiratory research》2011,12(1):57