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人类肿瘤坏死因子相关凋亡配体活性部位基因在毕赤酵母中的表达 被引量:3
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作者 徐红 赖心田 +2 位作者 叶凯 马辉文 洪葵 《生物工程学报》 CAS CSCD 北大核心 2003年第2期163-167,共5页
探讨了人类肿瘤坏死因子相关凋亡配体 (TRAIL)生物活性部分在巴斯德毕赤酵母 (Pichiapastoris)中的分泌表达。由于TRAIL活性部位区第 149 15 0氨基酸含有一个Kex2位点 ,根据Pichiapastoris分泌表达的特点 ,对 149位氨基酸进行了改造 ,... 探讨了人类肿瘤坏死因子相关凋亡配体 (TRAIL)生物活性部分在巴斯德毕赤酵母 (Pichiapastoris)中的分泌表达。由于TRAIL活性部位区第 149 15 0氨基酸含有一个Kex2位点 ,根据Pichiapastoris分泌表达的特点 ,对 149位氨基酸进行了改造 ,使其编码的氨基酸由Arg突变为Lys。这样使TRAIL在分泌的过程中不被Kex2切割 ,保证了片段的完整。序列分析正确后 ,将编码TRAIL的可溶区的基因片段插入到酵母表达载体pPIC9K中 ,使之位于α 因子信号肽下游 ,且与之同框 ,构建分泌型表达载体pPIC9K TRAIL。采用原生质体法将重组质粒转化Pichiapastoris菌株GS115 ,获基因工程菌株Pichiapastoris(pPIC9K TRAIL)。将工程菌用甲醇诱导培养 3~ 4d ,对摇瓶发酵的培养上清进行SDS PAGE、Westernblot分析和体外生物活性检测 ,结果发现发酵液中分子量约为 19kD和 38kD的蛋白质能被TRAIL多克隆抗体特异性识别 ,且具有在体外诱导肿瘤细胞凋亡的活性。通过薄层扫描分析显示目的蛋白最高可占可溶性总蛋白的 36 %。上述实验结果表明 ,在Pichiapastoris中表达的TRAIL能以寡聚体的形式存在并且具有生物学活性。 展开更多
关键词 表达 寡核苷酸介导 突变作用 巴斯德毕赤酵母 人类肿瘤坏死因子 凋亡配体
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人GITR基因的克隆和序列分析 被引量:4
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作者 仝佳 王胜军 +3 位作者 毛朝明 杨云良 陈君 杨敏 《江苏大学学报(医学版)》 CAS 2007年第1期15-18,共4页
目的:克隆人GITR基因全长编码区的cDNA,同时对其序列进行分析。方法:采用RT-PCR方法,从正常人外周血单个核细胞获得GITR基因的cDNA,克隆至pGEM-T载体,选择阳性克隆并进行序列测定。结果:扩增得到的人GITR基因编码区cDNA的全长726 bp,编... 目的:克隆人GITR基因全长编码区的cDNA,同时对其序列进行分析。方法:采用RT-PCR方法,从正常人外周血单个核细胞获得GITR基因的cDNA,克隆至pGEM-T载体,选择阳性克隆并进行序列测定。结果:扩增得到的人GITR基因编码区cDNA的全长726 bp,编码241个氨基酸残基,与GeneBank注册的序列完全一致。结论:获得人类GITR基因的克隆,为进一步研究其生物学功能奠定了基础。 展开更多
关键词 人类糖皮质激素诱导的肿瘤坏死因子受体 CDNA克隆 RT-PCR 调节性T细胞
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Pneumocystis jiroveci pneumonia and pneumomediastinum in an anti-TNFαnaive patient with ulcerative colitis
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作者 James C Lee Deborah C Bell +1 位作者 Richard M Guinness Tariq Ahmad 《World Journal of Gastroenterology》 SCIE CAS CSCD 2009年第15期1897-1900,共4页
We report the case of a 21-year-old man who was noted to have pneumomediastinum during an admission for an acute flare of ulcerative colitis. At that time, he was on maintenance treatment with azathioprine at a dose o... We report the case of a 21-year-old man who was noted to have pneumomediastinum during an admission for an acute flare of ulcerative colitis. At that time, he was on maintenance treatment with azathioprine at a dose of 2.25 mg/kg per day, and had not received supplementary steroids for 9 mo. He had never received anti-tumor necrosis factor (TNF)α therapy. Shortly after apparently effective treatment with intravenous steroids and an increased dose of azathioprine, he developed worsening colitic and new respiratory symptoms, and was diagnosed with Pneumocystis jiroveci (carinii) pneumonia (PCP). Pneumomediastinum is rare in immunocompetent hosts, but is a recognized complication of PCP in human immunodeficiency virus (HIV) patients, although our patient's HIV test was negative. Treatment of PCP with co-trimoxazole resulted in resolution of both respiratory and gastrointestinal symptoms, without the need to increase the steroid dose. There is increasing vigilance for opportunistic infections in patients with inflammatory bowel disease following the advent of anti-TNFα therapy. This case emphasizes the importance of considering the possibility of such infections in all patients with inflammatory bowel disease, irrespective of the immunosuppressants they receive, and highlights the potential of steroid-responsive opportunistic infections to mimic worsening colitic symptoms in patients with ulcerative colitis. 展开更多
关键词 Pneumocystis jiroveci Ulcerative colitis PNEUMOMEDIASTINUM AZATHIOPRINE CORTICOSTEROIDS
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Curative effect of Xuebijing injection on severe pulmonary contusion 被引量:6
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作者 Yi Chen Huasheng Tong +5 位作者 Wang Zhang Xingqin Zhang Zhigguo Pan Junming Qiu Ruihuan Pan Lei Su 《Journal of Traditional Chinese Medicine》 SCIE CAS CSCD 2013年第6期743-751,共9页
OBJECTIVE: To investigate the curative effects of Xuebijing (XBJ) injection, a Chinese patent medi- cine, on severe pulmonary contusion (PC). METHODS: Sixty-three patients with PC were ran- domized to convention... OBJECTIVE: To investigate the curative effects of Xuebijing (XBJ) injection, a Chinese patent medi- cine, on severe pulmonary contusion (PC). METHODS: Sixty-three patients with PC were ran- domized to conventional therapy plus XBJ injec- tion (n=33) or conventional therapy alone (n=30). Between groups differences in corticosteroid treat- ment, immune regulation therapy, hemofiltration, infusion volume, transfusion volume and antibiotic period were measured, as were intensive care unit(ICU)-free time, ventilation time, 28-day mortality rate and incidence of ventilation-associated pneu- monia (VAP). Serum concentrations of procalcito- nin (PCT), tumor necrosis factor-a (TNF-a), interleu- kin (IL)-6, and 11_-10, white blood cell (WBC) counts and percentages of human leukocyte antigen DR/ CD14+ (HLA-DR/CD14+) peripheral blood mononu- clear cells were compared. Markers of ventilation were determined by blood gas analysis and ventila- tor parameters. RESULTS: WBC counts and serum concentrations of PCT, TNF-a, 11.-6 and IL-10 were reduced signifi- cantly more quickly, and CD14+ percentage was in- creased significantly earlier, in the XBJ group than in the control group (P〈0.05 each). The level of ven- tilation and oxygenation index were ameliorated earlier in the XBJ than in the control group (P〈 0.05). XBJ treatment significantly reduced ICU-free time, ventilation time and incidence of VAP (P〈0.05 each), but had no effect on 28-day mortality rate (P〉0.05). CONCLUSION: XBJ treatment can shorten ICU-free and ventilation times and reduce the incidence of VAP, improving outcomes in patients with severe PC. XBJ may act by regulating inflammation and im- munity, alleviating systemic inflammatory response syndrome induced by trauma. 展开更多
关键词 CONTUSIONS Ventilator-induced lung in-jury Systemic inflammatory response syndrome Immunity HLA-DR antigens PROCALCITONIN Xuebi-jing injection
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