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Study of recombinant human interleukin-12 for treatment of complications after radiotherapy for tumor patients 被引量:7
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作者 Na Guo Wen-Qin Wang +8 位作者 Xiao-Jing Gong Lei Gao Li-Rong Yang Wei-Na Yu Hong-Yu Shen Ling-Qin Wan Xi-Feng Jia Yi-Shan Wang Yi Zhao 《World Journal of Clinical Oncology》 CAS 2017年第2期158-167,共10页
AIM To evaluate the treatment effects of recombinant human interleukin-12(rh IL-12) on radiotherapy complications, such as severe myelosuppression or pancytopenia, the decline or imbalance of immune function, etc.METH... AIM To evaluate the treatment effects of recombinant human interleukin-12(rh IL-12) on radiotherapy complications, such as severe myelosuppression or pancytopenia, the decline or imbalance of immune function, etc.METHODS The patients received high-dose and short-course precise radiotherapy, such as Cyber knife and image-guided radiotherapy(IGRT), which can cause myelosuppression or pancytopenia and immune function decline within a short time. One-hundred subjects were enrolled in the study, and 50 were randomized to a treatment group which used rh IL-12 and 50 were randomized to a control group which used symptomatic and supportive therapy after radiotherapy. The 50 subjects in the treatment group were further divided into five subgroups and intervenedwith rh IL-12 at a dose of 50, 100, 150, 200 or 250 ng/kg respectively. The dose-effect relationship was observed. RESULTS Rh IL-12 significantly attenuated the decrease of peripheral blood cells in the treatment group, and immune function was improved after treatment. Due to the different radiation doses, there was a fluctuation within 12 h after treatment but mostly showing an increasing trend. As to the clinical manifestations, 2 patients in the 250 ng/kg subgroup showed low fever after administration, 1 patient in the 200 ng/kg subgroup and 2 patients in the 250 ng/kg subgroup showed mild impairment of liver function during the observation period.CONCLUSION Rh IL-12 has effective therapeutic and protective effects on complications following radiotherapy, such as the decline of blood cells, myelosuppression and the decline or imbalance of immune function, which indicated good prospects for development and application. 展开更多
关键词 recombinant human interleukin-12 Cancer PREVENTION RADIOTHERAPY COMPLICATIONS Clinical research
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Recombinant human interleukin-11 for treatment of chemotherapy-induced thrombocytopenia in patients with gastrointestinal cancer 被引量:1
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作者 Jie Li Lin Shen Yan Li Xiaodong Zhang Jian Li Jifang Gong Wei Deng 《The Chinese-German Journal of Clinical Oncology》 CAS 2007年第5期450-452,共3页
Objective: To evaluate the efficacy and safety of recombinant human interleukin-11 (rhIL-11) for the chemotherapy-induced thrombocytopenia in patients with gastrointestinal cancer. Methods: It was an opened and no... Objective: To evaluate the efficacy and safety of recombinant human interleukin-11 (rhIL-11) for the chemotherapy-induced thrombocytopenia in patients with gastrointestinal cancer. Methods: It was an opened and non-randomized controlled clinical study. When the platelet counts was under 75 × 10^9/L after chemotherapy, rhlL-11 was administered 25 μg/(kg·d) as a daily SC injection last for 7-14 days, or discontinued when platelet counts 〉 100 × 10^9/L. Results: Seventysix patients were enrolled into this study. The treatment group and the control group had thirty-eight cases, respectively. The mean recovery time to PLT ≥ 100 × 10^9/L was 8.1 days in treatment group, while in control group was 12.2 days (P 〈 0.01). Moreover, the mean recovery time from PLT 〈_ 50 × 10^9/L to 〉 100 × 10^9/L was 8.9 days in treatment group, while in control group was 12.9 days (P 〈 0.05). There was a statistical difference between the two groups. Major side effects included edema, fever, articular muscle soreness, but they were all mild and well tolerable. Conclusion: rhIL-11 can be safely and effectively used for the treatment of chemotherapy-induced thrombocytopenia in patients with gastrointestinal cancer. 展开更多
关键词 recombinant human interleukin-11 (rhIL-11) gastrointestinal cancer thrembocytopenia CHEMOTHERAPY
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Neuronal changes in the retinal ganglion cell layer following recombinant human interleukin-2 intravitreal injection in a rat model of chronically elevated intraocular pressure
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作者 Ning Li Jing Wang Xuan Zou Juanlian Cui Xuanchu Duan 《Neural Regeneration Research》 SCIE CAS CSCD 2010年第24期1888-1894,共7页
Intraperitoneal injection of recombinant human interleukin-2(rhIL-2)inhibits neuronal apoptosis in the chronic ocular hypertension retinal ganglion cell layer.Intravitreous injection was performed on retinal ganglio... Intraperitoneal injection of recombinant human interleukin-2(rhIL-2)inhibits neuronal apoptosis in the chronic ocular hypertension retinal ganglion cell layer.Intravitreous injection was performed on retinal ganglion cells in a Wistar rat model of chronically elevated intraocular pressure to observe the effects of LY294002 and AG490 on retinal ganglion cell survival,macrophage activation,and PI3K/Akt and JAK/STAT activation.The number of retinal ganglion cells in the rhIL-2 treatment group was much greater than in the normal control and phosphate-buffered saline groups.Western blot analysis revealed low Akt and STAT3 protein expression in the retina after 3-hour intravitreous injections of rhIL-2.However,protein expression was increased at 12 hours,but decreased again at 24 hours,with very low expression at 96 hours.LY294002 and AG490,which are inhibitors of the PI3K/Akt and JAK/STAT3 signal pathways,prevented upregulation of Akt and STAT3 protein expression in the retina,respectively.Intravitreous injection of rhIL-2 exhibited neuroprotective effects by decreasing retinal ganglion cell layer damage in a rat model of chronic glaucoma.These results suggest that intravitreal injection of rhIL-2 could induce the PI3K/Akt and JAK/STAT3 signaling pathways to protect retinal ganglion cells in chronically elevated intraocular pressure models. 展开更多
关键词 GLAUCOMA NEUROPROTECTION signal pathway recombinant human interleukin-2 retinal ganglion cells
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应用重组PCR技术构建人单链白细胞介素12融合基因 被引量:11
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作者 张梅 司履生 王一理 《免疫学杂志》 CAS CSCD 北大核心 2001年第1期22-26,共5页
目的构建人单链白细胞介素 12 (hsc IL- 12 )融合基因并在真核细胞中进行表达 ,为进一步研究 hsc IL- 12融合蛋白的生物学活性奠定基础。方法应用重组 PCR技术将 h IL - 12 p40和 p35亚单位两段编码序列的 c DNA通过一疏水性多肽接头 (G... 目的构建人单链白细胞介素 12 (hsc IL- 12 )融合基因并在真核细胞中进行表达 ,为进一步研究 hsc IL- 12融合蛋白的生物学活性奠定基础。方法应用重组 PCR技术将 h IL - 12 p40和 p35亚单位两段编码序列的 c DNA通过一疏水性多肽接头 (Gly4Ser) 3碱基序列进行前后拼接 (IL- 12 p40 -多肽接头 - p35 ) ,构建 hsc IL- 12融合基因 ,并进行核苷酸序列测定 ,进一步将 hsc IL - 12融合基因插入 pc DNA3.1真核表达载体中 ,转染 COS- 7细胞表达 hsc IL - 12融合蛋白 ,并行 Western blot分析。结果该融合基因经 DNA序列分析表明 :p40、p35、linker的连接顺序、方向及序列完全正确 ,融合基因可在 COS- 7细胞中表达hsc IL- 12融合蛋白。Western blot显示该融合蛋白分子量为 70 0 0 0 u,可与鼠抗人 IL- 12 p40 /p70单克隆抗体特异性结合。结论重组 PCR技术是十分有效。 展开更多
关键词 重组PCR 人白细胞介素12 人单链白细胞介素 融合基因
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rhIL-12对恒河猴造血系统辐射防护作用的初步研究 被引量:12
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作者 熊国林 赵毅 +10 位作者 邢爽 申星 宁学成 卢世香 李建 郭玲玲 郝锐 陈廷超 缪锦来 贺济琛 罗庆良 《中国实验血液学杂志》 CAS CSCD 北大核心 2013年第1期150-154,共5页
本研究旨在初步观察重组人白介素-12(rhIL-12)对急性辐射损伤猴造血系统的防护和治疗作用,为同类型病人的临床救治提供实验依据。在体外实验中,观察了不同浓度rhIL-12(0、1、5、25、125和625 ng/ml)对来自人和健康成年猴骨髓造血祖细胞... 本研究旨在初步观察重组人白介素-12(rhIL-12)对急性辐射损伤猴造血系统的防护和治疗作用,为同类型病人的临床救治提供实验依据。在体外实验中,观察了不同浓度rhIL-12(0、1、5、25、125和625 ng/ml)对来自人和健康成年猴骨髓造血祖细胞集落形成能力的影响。在体内实验中,用11只经3.0 Gy全身照射后90 d活存猴再经致死剂量60Coγ(6.0 Gy)射线全身1次照射以建立重度骨髓型急性放射病动物模型。模型动物被随机分为照射对照组(n=4)、单次给药组(n=3)和分次给药组(n=4)。单次给药组于照射后2 h皮下注射rhIL-12 4μg/kg,分次给药组分别于照射后2 h和3、6、9 d皮下注射rhIL-12 1μg/kg,对照组于皮下注射同样体积的PBS。观察照射后动物活存情况和外周血象变化。体外培养结果显示,不同浓度rhIL-12可明显促进正常猴和人骨髓单个核细胞形成各系造血祖细胞集落,特别是CFU-E和CFU-GM最为明显。照射对照组动物于照射后22 d内全部死亡,死亡动物平均活存时间为(20.3±1.2)d;rhIL-12单次给药组3只动物全部存活,rhIL-12分次给药组1只动物于照射后17 d因贫血死亡。与对照组比较,rhIL-12治疗能明显提高动物存活(P=0.018),并促进外周血白细胞数、血红蛋白水平、血小板以及网织红细胞数的恢复(P<0.05)。结论:rhIL-12可明显促进灵长类骨髓造血祖细胞体外集落形成,并明显促进重度骨髓型急性放射病猴的造血功能恢复,提高动物存活率。 展开更多
关键词 重组人白细胞介素-12 辐射损伤 急性放射病 恒河猴
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人IL-12/慢病毒重组体对结肠癌干细胞“干性”的影响 被引量:3
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作者 尹晓玲 王正中 +3 位作者 赵银晶 陈华俊 杨晓琼 梁后杰 《中国免疫学杂志》 CAS CSCD 北大核心 2013年第2期161-167,共7页
目的:构建并鉴定人白介素12(Interleukin-12,IL-12)-慢病毒过表达重组体,观察其在结肠癌干细胞(Cancerstem cell,CSCs)中的表达及其作用,为后续动物试验做准备。方法:PCR扩增pORF-hIL-12质粒获得IL-12基因,通过双酶切及连接后构建IL-12... 目的:构建并鉴定人白介素12(Interleukin-12,IL-12)-慢病毒过表达重组体,观察其在结肠癌干细胞(Cancerstem cell,CSCs)中的表达及其作用,为后续动物试验做准备。方法:PCR扩增pORF-hIL-12质粒获得IL-12基因,通过双酶切及连接后构建IL-12/慢病毒过表达重组体;分离鉴定结肠CSCs,将IL-12/慢病毒过表达重组体转染结肠CSCs建立IL-12过表达模型,检测IL-12在结肠CSCs的表达及对结肠CSCs"干性"的影响。结果:PCR、酶切鉴定及测序证实IL-12/慢病毒过表达重组体构建正确,流式细胞仪成功分选出CD133+CD44+SW480结肠CSCs,IL-12/慢病毒转染结肠CSCs后,RT-PCR及West-ern blot证实结肠CSCs培养上清均有IL-12表达,IL-12抑制结肠CSCs自我更新,促进其分化。结论:IL-12/慢病毒过表达重组体能在结肠CSCs中稳定表达并能抑制结肠CSCs自我更新及分化。 展开更多
关键词 人白介素12重组体 构建 癌症干细胞 表达 干性
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重组人IL-12真核表达载体的基因佐剂功能 被引量:2
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作者 王丽娜 孙运芳 +4 位作者 吕锐 于红 曹永献 刘志军 张文卿 《中国生物制品学杂志》 CAS CSCD 2008年第7期553-556,共4页
目的探讨重组人IL-12真核表达载体(pcDNA6-p70)对核酸疫苗(HCMV pp65基因重组腺病毒,Adeno-pp65)的基因佐剂功能。方法pcDNA6-p70与Adeno-pp65共免疫BALB/c小鼠,每2周1次,共4次,同时设立Adeno-pp65对照组及生理盐水对照组。于初次免疫后... 目的探讨重组人IL-12真核表达载体(pcDNA6-p70)对核酸疫苗(HCMV pp65基因重组腺病毒,Adeno-pp65)的基因佐剂功能。方法pcDNA6-p70与Adeno-pp65共免疫BALB/c小鼠,每2周1次,共4次,同时设立Adeno-pp65对照组及生理盐水对照组。于初次免疫后第6周末尾静脉取血,第9周处死小鼠,分离脾细胞,分别检测细胞内细胞因子水平、CTL杀伤活性、特异性淋巴细胞增生和IFN-γ水平,并观察pcDNA6-p70对小鼠的安全性。结果共免疫组HCMV特异性CD4/IFN-γ、CD8/IFN-γ双标记阳性细胞的百分率均高于Adeno-pp65组及生理盐水对照组,差异均有显著意义;其CTL杀伤活性、HCMV特异性淋巴细胞增生水平及脾淋巴细胞特异性IFN-γ释放水平均高于Adeno-pp65组和生理盐水对照组,差异均有显著意义。pcDNA6-p70肌肉注射昆明小鼠,其体温、体重及一般情况与对照组差异均无显著意义。结论pcDNA6-p70与Adeno-pp65共免疫可提高核酸疫苗的免疫效果,且具有较好的安全性。 展开更多
关键词 白细胞介素-12 基因佐剂 人巨细胞病毒 pp65基因 重组腺病毒
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重组人单链白细胞介素12融合基因(rhscIL-12)的构建和真核表达 被引量:1
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作者 张梅 司履生 +1 位作者 王一理 耿宜苹 《西安医科大学学报》 CSCD 2000年第4期297-302,共6页
克隆人 IL - 1 2 ( h IL - 1 2 ) p40和 p35亚单位 c DNA,构建人单链 IL- 1 2 ( rhsc IL- 1 2 )融合基因 ,并在哺乳动物细胞中进行表达。方法 从经 PDBu刺激的 EBV转化的人 B淋巴母细胞株 NC37中提取 m RNA,经 RT- PCR分别获得 h IL- 1... 克隆人 IL - 1 2 ( h IL - 1 2 ) p40和 p35亚单位 c DNA,构建人单链 IL- 1 2 ( rhsc IL- 1 2 )融合基因 ,并在哺乳动物细胞中进行表达。方法 从经 PDBu刺激的 EBV转化的人 B淋巴母细胞株 NC37中提取 m RNA,经 RT- PCR分别获得 h IL- 1 2 p40和 p35亚单位编码序列的 c DNA,运用重组 PCR技术将两段基因通过一疏水性多肽接头 ( Gly4 Ser) 3 DNA序列进行体外基因重组 ,构建 rhsc IL- 1 2融合基因 ,将其插入 pc DNA3.1 ( + )真核表达载体 ,经脂质体转染 COS7细胞进行表达 ,Western blot进行分析。结果 所克隆的 h IL- 1 2 p40、p35c DNA序列和构建的 rhsc IL - 1 2融合基因 DNA序列均经测序得以证实 ,融合基因可在 COS7中表达其产物 rhsc IL- 1 2融合蛋白 ,其分子量为 70 KD,可与鼠抗人 IL - 1 2 p40 /p70单克隆抗体特异性结合。结论 本研究结果为进一步探讨 rhsc IL- 1 2融合蛋白的生物学活性和特性奠定了基础 ,也为 rhsc IL- 1 展开更多
关键词 重组人单链白细胞介素12 融合基因 真核表达
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肿瘤治疗性多肽疫苗——重组人纽表位肽12质控方法的研究
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作者 纪宏 王军志 +2 位作者 饶春明 张翊 王旻 《药学学报》 CAS CSCD 北大核心 2004年第5期359-362,共4页
目的 建立重组人纽表位肽12质控方法和检定规程。方法 采用FVB/N转neu基因小鼠,以ELISA法检测给药后小鼠的抗体阳转百分率评价其生物学活性,用胃蛋白酶酶切分析肽图,用SEC-HPLC法测定抗原含量。结果与结论 建立了重组人纽表位肽12的生... 目的 建立重组人纽表位肽12质控方法和检定规程。方法 采用FVB/N转neu基因小鼠,以ELISA法检测给药后小鼠的抗体阳转百分率评价其生物学活性,用胃蛋白酶酶切分析肽图,用SEC-HPLC法测定抗原含量。结果与结论 建立了重组人纽表位肽12的生物学活性、肽图分析、抗原含量测定等质控方法及检定规程,并已用于重组人纽表位肽12产品的检定。 展开更多
关键词 肿瘤 治疗 多肽疫苗 重组人纽表位肽12 质量控制
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抗重组人白细胞介素-12单克隆抗体的制备及初步应用
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作者 李俐 马颖哲 +1 位作者 张家颖 张桂荣 《中国生物制品学杂志》 CAS CSCD 2008年第8期705-707,共3页
目的制备抗重组人白细胞介素-12(rhIL-12)单克隆抗体(McAb),并建立检测IL-12的双抗体夹心ELISA方法。方法以纯化的rhIL-12(p70)免疫BALB/c小鼠,制备单克隆抗体,建立双抗体夹心ELISA法,检测胃癌患者术前、术后血清IL-12水平的变化。结果... 目的制备抗重组人白细胞介素-12(rhIL-12)单克隆抗体(McAb),并建立检测IL-12的双抗体夹心ELISA方法。方法以纯化的rhIL-12(p70)免疫BALB/c小鼠,制备单克隆抗体,建立双抗体夹心ELISA法,检测胃癌患者术前、术后血清IL-12水平的变化。结果筛选出3株稳定分泌抗rhIL-12的单抗杂交瘤细胞株,纯化后单抗纯度达95%以上。免疫球蛋白亚类2株为IgG1,1株为IgG2a,轻链属!型,相对亲和力依次为EM5>EM6>EV9。Western blot及ELISA交叉试验表明,单抗具有良好的特异性。建立的双抗体夹心ELISA方法检测rhIL-12的最低检出限为20pg/ml,线性范围为25~3200pg/ml。应用此法检测胃癌患者术前血清IL-12水平较正常对照组明显降低,术后10d明显升高。术后给予免疫增强型NE组IL-12水平较未给予组明显升高。结论制备的抗rhIL-12单克隆抗体,能够用于IL-12的体内外免疫学检测,为IL-12的研究、检测及临床应用奠定了基础。 展开更多
关键词 重组人白细胞介素-12 单克隆抗体 双抗体夹心ELISA 胃癌
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重组人白介素12质量标准与检测方法研究
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作者 王威 李永红 +2 位作者 饶春明 杭太俊 王军志 《药物分析杂志》 CAS CSCD 北大核心 2006年第3期287-290,共4页
目的:建立白介素12的质量标准与检测方法,用于该产品的质量控制。方法:采用诱导 PBMC 生成 IFN-γ的方法测定生物学活性;用胰酶酶解分析肽图;用分子排阻 HPLC、离子交换 HPLC 和 SDS-PAGE 法分析蛋白纯度;免疫印迹及 N 端氨基酸序列测... 目的:建立白介素12的质量标准与检测方法,用于该产品的质量控制。方法:采用诱导 PBMC 生成 IFN-γ的方法测定生物学活性;用胰酶酶解分析肽图;用分子排阻 HPLC、离子交换 HPLC 和 SDS-PAGE 法分析蛋白纯度;免疫印迹及 N 端氨基酸序列测定分析、鉴定产品特性。按照《中国药典》三部2005年版的有关规定进行了其他项目检查。结果:用建立的方法对重组人白介素12成品及原液进行检定,结果各项指标均符合《中国药典》的要求。结论:建立了重组人白介素12的质量标准,并可有效地控制制品的质量。 展开更多
关键词 重组人白介素12 质量标准 生物学活性
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重组人纽表位肽12生物学活性测定方法的研究
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作者 纪宏 王军志 +1 位作者 饶春明 张翊 《中国肿瘤生物治疗杂志》 CAS CSCD 2003年第4期280-284,共5页
目的:建立适用于重组人纽表位肽12体外质量控制的生物学活性测定方法,用于该制品的生物学活性评价。方法:采用小鼠,比较不同品系、给药浓度、给药周期等条件下应用ELISA方法检测出的小鼠相应抗体水平,确定最佳实验条件,以建立相应的体... 目的:建立适用于重组人纽表位肽12体外质量控制的生物学活性测定方法,用于该制品的生物学活性评价。方法:采用小鼠,比较不同品系、给药浓度、给药周期等条件下应用ELISA方法检测出的小鼠相应抗体水平,确定最佳实验条件,以建立相应的体外活性测定方法。结果:FVB/N转neu基因小鼠(TgN MMTVneu 202 Mul,Jackson Lab.,USA)对重组人纽表位肽12的反应存在量——效关系,并确定了最佳测定条件和给药剂量,用抗体阳转百分率作为评价指标,样品测定重复性较好,CV%值为6.7%(n=4),结果可靠。结论:已建立的FVB/N转neu基因小鼠测定重组人纽表位肽12生物学活性的方法可用于重组人纽表位肽12生物学活性的常规评价。 展开更多
关键词 重组人纽表位肽12 生物学活性 FVB/N转neu基因小鼠 ELISA
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重组人纽表位肽12的胃蛋白酶切肽图分析方法的研究
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作者 纪宏 王军志 王旻 《药物生物技术》 CAS CSCD 2004年第5期290-293,共4页
采用反相高效液相色谱法建立重组人纽表位肽12的标准肽图分析法,固定相为十八烷基硅烷键合硅胶,流动相A液0.1%TFA的水溶液,流动相B液0.1%TFA的乙腈溶液;梯度洗脱程序0~70min内B%由0%增至70%。结果3批重组人纽表位肽12样品的反相高效液... 采用反相高效液相色谱法建立重组人纽表位肽12的标准肽图分析法,固定相为十八烷基硅烷键合硅胶,流动相A液0.1%TFA的水溶液,流动相B液0.1%TFA的乙腈溶液;梯度洗脱程序0~70min内B%由0%增至70%。结果3批重组人纽表位肽12样品的反相高效液相色谱图完全一致,与重组人纽表位肽12理化对照品的肽图比较,完全吻合,同时胃蛋白酶并不干扰肽图的测定,该法准确度高、重现性好,可用于重组人纽表位肽12的质量控制。 展开更多
关键词 重组人纽表位肽12 肽图 反相高效液相色谱法 胃蛋白酶切
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rhIL-12联合HBsAg刺激健康人外周血单核细胞对IL-12R的影响
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作者 李茹冰 李若冰 +3 位作者 王翠玲 张宜俊 李超 傅泳航 《中国生化药物杂志》 CAS CSCD 北大核心 2012年第5期524-527,共4页
目的确认重组人白细胞介素12(rhIL-12)联合乙型肝炎表面抗原(HBsAg)可增强健康人外周血单核细胞(PBMC)产生特异性细胞免疫应答,并分析其细胞来源,及对IL-12R的影响,探讨hIL-12联合HBsAg作为治疗型乙型肝炎疫苗的应用前景。方法 8名健康... 目的确认重组人白细胞介素12(rhIL-12)联合乙型肝炎表面抗原(HBsAg)可增强健康人外周血单核细胞(PBMC)产生特异性细胞免疫应答,并分析其细胞来源,及对IL-12R的影响,探讨hIL-12联合HBsAg作为治疗型乙型肝炎疫苗的应用前景。方法 8名健康人PBMC分别与rhIL-12(1 ng/mL)、rHBsAg(500 ng/mL)和rhIL-12(1ng/mL)+rHBsAg(500 ng/mL)体外孵育,采用流式细胞术检测CD4+IFN-γ(γ-干扰素)+T细胞、CD8+IFN-γ+T细胞及CD56+IFN-γ+NK细胞百分比,并测定表达IL-12Rβ1、IL-12Rβ2的细胞百分比。结果 rhIL-12与rHBsAg联合组CD8+IFN-γ+T细胞及CD56+IFN-γ+NK细胞百分比均较对照组明显升高,且有非常显著差异(P<0.01),联合组CD8+IL-12Rβ1和CD56+IL-12Rβ1与单用rhIL-12或rHBsAg组比较均显著升高。结论提示rhIL-12联合rHBsAg诱生的IFN-γ主要来自CD56+NK细胞、CD8+T细胞,其次是CD4+T细胞;rhIL-12和rHBsAg单独均能上调健康人PBMCs表面IL-12R的表达,rhIL-12与rHBsAg联合具有增强作用。 展开更多
关键词 乙型肝炎表面抗原 重组人白细胞介素12 外周血单个核细胞 Γ干扰素 IL-12R
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重组人白细胞介素12对健康人外周血单个核细胞产生γ干扰素影响的研究 被引量:1
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作者 段祥 王翠玲 +7 位作者 王增松 谢飞群 揭燕 夏书奇 曾振飞 李茹冰 傅泳航 张宜俊 《检验医学》 CAS 北大核心 2009年第8期586-590,共5页
目的建立重组人白细胞介素12(rhIL-12)的生物活性检测方法,观察rhIL-12诱导健康人外周血单个核细胞(PBMC)产生γ干扰素(IFN-γ)的水平与剂量和时间之间的关系,揭示其变化规律。方法rhIL-12供试品刺激健康人PBMC分泌IFN-γ,经国际标准品... 目的建立重组人白细胞介素12(rhIL-12)的生物活性检测方法,观察rhIL-12诱导健康人外周血单个核细胞(PBMC)产生γ干扰素(IFN-γ)的水平与剂量和时间之间的关系,揭示其变化规律。方法rhIL-12供试品刺激健康人PBMC分泌IFN-γ,经国际标准品校正,建立rhIL-12生物活性测定方法。以1 ng/mL浓度rhIL-12诱导健康人PBMC,分别于24、48、72、96和120 h 5个时间点用酶联免疫吸附试验(ELISA)测定培养上清的IFN-γ水平。测定0.1、0.5和2.5 ng/mL 3个浓度rhIL-12诱导人PBMC 72 h后培养上清的IFN-γ水平,用Prism 5.0统计软件的重复测量方差分析法对所得数据进行统计学处理。结果rhIL-12供试品的生物活性曲线呈典型S型,IFN-γ含量为最大浓度一半时的供试品浓度(ED50)为(1.008±0.238)ng/mL,供试品rhIL-12效价为7 938 IU/mL。测定1ng/mL浓度rhIL-12刺激健康人PBMC后5个时间点培养上清中IFN-γ显示,不同个体高峰出现时间有所不同,在高峰出现前均呈现递增趋势。高、中、低3个浓度rhIL-12分别诱导健康人PBMC产生的IFN-γ水平与浓度之间呈显著量效关系,且在0.1 ng/mL的极低浓度即可诱生高水平IFN-γ。结论rhIL-12供试品生物活性良好,与国际标准品比对基本一致,不同人群对rhIL-12的应答高峰存在个体差异,以诱生IFN-γ作为评价rhIL-12生物活性的指标具有明显的量效关系。 展开更多
关键词 重组人白细胞介素12 Γ干扰素 细胞免疫
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HBsAg联合rhIL-12对慢性乙型肝炎患者PBMC诱生IFN-γ的作用 被引量:1
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作者 王翠玲 李茹冰 +7 位作者 王增松 夏书奇 段祥 谢飞群 曾振飞 揭燕 傅泳航 张宜俊 《检验医学》 CAS 北大核心 2009年第9期671-675,共5页
目的确认重组人白细胞介素12(rhIL-12)联合乙型肝炎表面抗原(HBsAg)可增强慢性乙型肝炎患者外周血单个核细胞(PBMC)对HBsAg产生特异性细胞免疫应答,探讨rhIL-12联合HBsAg作为治疗型乙型肝炎疫苗的应用前景。方法取8名正常人和16例慢性... 目的确认重组人白细胞介素12(rhIL-12)联合乙型肝炎表面抗原(HBsAg)可增强慢性乙型肝炎患者外周血单个核细胞(PBMC)对HBsAg产生特异性细胞免疫应答,探讨rhIL-12联合HBsAg作为治疗型乙型肝炎疫苗的应用前景。方法取8名正常人和16例慢性乙型肝炎患者的PBMC,分别用不同浓度的HBsAg、rhIL-12单独体外刺激,再用HBsAg联合rhIL-12(0.01、0.1和1.0ng/mL)刺激患者PBMC,72h后用酶联免疫吸附试验(ELISA)测定培养上清中的γ干扰素(IFN-γ)。结果HBsAg或rhIL-12单独刺激仅产生低水平的IFN-γ,联合使用HBsAg和rhIL-12刺激后,患者PBMC产生的IFN-γ平均水平显著高于单独使用HBsAg或rhIL-12(P均<0.05)。HBsAg(0.5μg/mL)联合rhIL-12(0.01、0.1和1.0ng/mL)刺激患者PBMC产生的IFN-γ与rhIL-12的浓度之间呈明显的量效关系。结论HBsAg和rhIL-12联合刺激乙型肝炎患者PBMC,通过产生高水平IFN-γ激活乙型肝炎患者特异性细胞免疫。 展开更多
关键词 乙型肝炎表面抗原 重组人白细胞介素12 外周血单个核细胞 细胞免疫 Γ干扰素
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Why interleukin-10 supplementation does not work in Crohn's disease patients 被引量:9
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作者 Gareth J Marlow Dominique van Gent Lynnette R Ferguson 《World Journal of Gastroenterology》 SCIE CAS 2013年第25期3931-3941,共11页
Inflammatory bowel diseases (IBD) such as Crohn's disease (CD) or ulcerative colitis are chronic intestina disorders, which are on the increase in "Westernised" countries. IBD can be caused by both genet... Inflammatory bowel diseases (IBD) such as Crohn's disease (CD) or ulcerative colitis are chronic intestina disorders, which are on the increase in "Westernised" countries. IBD can be caused by both genetic and environmental factors. Interleukin-10 (IL-10) is an immunoregulatory cytokine that has been identified as being involved in several diseases including IBD. Studies have shown that polymorphisms in the promoter region reduce serum levels of IL-10 and this reduction has been associated with some forms of IBD. Mouse models have shown promising results with IL-10 supplementation, as such IL-10 supplementation has been touted as being a possible alternative treatment for CD in humans. Clinical trials have shown that recombinant human IL-10 is safe and well tolerated up to a dose o 8 μg/kg. However, to date, the results of the clinica trials have been disappointing. Although CD activity was reduced as measured by the CD activity index IL-10 supplementation did not result in significantly reduced remission rates or clinical improvements when compared to placebo. This review discusses why IL-10supplementation is not effective in CD patients currently and what can be addressed to potentially make IL-10 supplementation a more viable treatment option in the future. Based on the current research we conclude that IL-10 supplementation is not a one size fits all treatment and if the correct population of patients is chosen then IL-10 supplementation could be of benefit. 展开更多
关键词 INFLAMMATORY BOWEL DISEASE Crohn’s DISEASE interleukin-10 recombinant human interleukin-10
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A multiple-dose pharmacokinetics of polyethylene glycol recombinant human interleukin-6 (PEG-rhIL-6) in rats 被引量:1
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作者 Xue-ling HE Hai-lin YIN +7 位作者 Jiang WU Ke ZHANG Yan LIU Tao YUAN Hai-lin RAO Liang LI Guang YANG Xue-mei ZHANG 《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 SCIE CAS CSCD 2011年第1期32-39,共8页
Radiation therapy has been widely applied in cancer treatment.However,it often causes thrombocytopenia (deficiency of white blood cells) as an adverse effect.Recombinant human interleukin-6 (rhIL-6) has been found to ... Radiation therapy has been widely applied in cancer treatment.However,it often causes thrombocytopenia (deficiency of white blood cells) as an adverse effect.Recombinant human interleukin-6 (rhIL-6) has been found to be a very effective way against this thrombocytopenia,but IL-6 has low stability in blood,which reduces its efficacy.To increases the stability and half-life of rhIL-6,it was modified by polyethylene glycol (PEG).The pharmacokinetics and the tissue distribution of PEG-rhIL-6 labeled with 125I were examined after subcutaneous injection in rats.The pharmacokinetic pattern of PEG-rhIL-6 was defined with linear-kinetics,and we fitted a one-compartment model with half-lives of 10.44–11.37 h (absorption,t1/2Ka) and 19.77–21.53 h (elimination,t1/2Ke),and peak concentrations at 20.51–21.96 h (tpeak) in rats.Half-lives and tpeak of PEG-rhIL-6 were longer than those of rhIL-6 previously reported.In the present study,for deposition of PEG-rhIL-6 in rats,the tissue distribution examination showed that blood was the major organ involved,rather than liver.However,as to the elimination of PEG-rhIL-6,the major organ was the kidney.The excretion fraction of the injection dose recovered from urine was 23.32% at 192 h after subcutaneous administration.Less than 6% of PEG-rhIL-6 was eliminated via the feces at 192 h.These results indicate that PEG-rhIL-6 is a good candidate drug formulation for patients with cancer. 展开更多
关键词 Polyethylene glycol recombinant human interleukin-6 PHARMACOKINETICS RAT
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Pharmacokinetics of recombinant human interleukin-3 in rhesus monkeys
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作者 汤仲明 刘秀文 屠敏 《Science China(Life Sciences)》 SCIE CAS 1997年第5期546-553,共8页
Concentration-time profiles of ^(125)I-labeled recombinant human interleukin-3 (125IrhIL-3) were de-termined by reverse phase high performance liquid chromatography (RHPLC) after intravenous and subcutaneous ad-minist... Concentration-time profiles of ^(125)I-labeled recombinant human interleukin-3 (125IrhIL-3) were de-termined by reverse phase high performance liquid chromatography (RHPLC) after intravenous and subcutaneous ad-ministration of the drug in 16 rhesus monkeys. The initial and terminal T1/2 in plasma after intravenous of 30 μg/kg were (0. 15 ±0.13) and (2. 21 ± 0. 59) h, respectively. Terminal half-lives after 30, 90 and 180μg/kg subcutaneous (s.c.) injections were 2. 0-3.8 h. Area under concentration-time curves (AUC) following s. c. were roughly in-creased with dose, while CL5 were similar among different dosages. The absorption rates were dependent on concentra-tion at injected site. Bioavailability was about 0.7 after s.c. Rapid biodegradation was found in plasma. Distribution profiles of total radioactivity were as follows: the highest level was found in urinary system; levels in bile-enteric sys-tem, lymph nodes, bone marrow and spleen were near to that in plasma, and level in brain was the lowest. The RH-PLC analysis revealed that kidney was one of the major organs for biodegradation. 展开更多
关键词 recombinant human interleukin-3 rhesus monkey PHARMACOKINETICS ^(125)l-labeling high perfor-mace liquid chromatography.
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Recombinant human heparin-binding neurite-promoting factor expressed with yeast stimulates neurites outgrowth
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作者 王毅超 陈峥嵘 +4 位作者 陈中伟 官孝群 宋后燕 吴欣 刘银坤 《Chinese Medical Journal》 SCIE CAS CSCD 2002年第9期1352-1357,150-151,共6页
OBJECTIVES: Heparin-binding neurite-promoting factor (HBNF) is a heparin-binding protein primarily found in the brain, which can stimulate neurite outgrowth in vitro. We expressed recombinant human heparin-binding neu... OBJECTIVES: Heparin-binding neurite-promoting factor (HBNF) is a heparin-binding protein primarily found in the brain, which can stimulate neurite outgrowth in vitro. We expressed recombinant human heparin-binding neurite-promoting factor (hrHBNF) using a yeast system, and observed its activity in stimulating neurite outgrowth in vitro. METHODS: cDNA encoding mature human HBNF was amplified from total RNA isolated from an 18-week aborted human fetal brain by RT-PCR method. After amplification, the HBNF cDNA gene was cloned into pPIC9K, a shuttle expression vector for yeast system. The positive clone of expression vector bearing HBNF cDNA gene was obtained by screening. Verified recombinant vector was then used to transform Pichia strain GS115 by electroporation. His(+) transformants were selected on minimal dextrose medium (MD) plates which were histidine free. His(+) yeast recombinants with multi-copy inserts were screened in vivo by their resistance to G418. PCR analysis was used to confirm the integration of the HBNF cDNA gene into the Pichia genome. Secreted expression of hrHBNF protein in culture medium was obtained when the positive clone containing the HBNF cDNA gene was induced by methanol. The hrHBNF product purified by gel chromatography was added to cultured rat pheochromocytoma (PC12) cells to observe its ability to stimulate neurite outgrowth. RESULTS: In the recombinant expression vector, the insert was sequenced to show exactly the sequence encoding human HBNF according to Genbank data. The HBNF cDNA gene was cloned downstream to the alpha-factor, and its open reading frame was in frame with the alpha-factor signal sequence in pPIC9K. SDS-PAGE showed that the molecular weight of the induced expression product was about 18 kDa, consistent with that of human HBNF reported in the literature. The protein product did promote neurite outgrowth in cultured rat pheochromocytoma (PC12) cells. CONCLUSION: Recombinant human heparin-binding neurite-promoting factor can be expressed with a yeast system, and its product possesses the biological activity to promote neurite outgrowth. 展开更多
关键词 Animals Base Sequence Carrier Proteins Cytokines DNA Complementary Genetic Vectors humans Molecular Sequence Data NEURITES PC12 Cells PICHIA Rats recombinant Proteins Research Support Non-U.S. Gov't Reverse Transcriptase Polymerase Chain Reaction
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