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
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
OBJECTIVE: To identify a gene engineering antibody against cystic echinococcosis in liver. METHODS: A single chain of variable fragment of human antibodies (ScFvs) was selected from the library by using affinity selec...OBJECTIVE: To identify a gene engineering antibody against cystic echinococcosis in liver. METHODS: A single chain of variable fragment of human antibodies (ScFvs) was selected from the library by using affinity selection technique with the recombinant antigen on solid surface. The positive clones were demonstrated by ELISA and their DNA sequences were also determined. RESULTS: The DNA sequence data showed that the antibody gene is composed of 768bp. In addition, a specific combination capacity with recombinant Echinococcus granulosus antigen B (r-EgB) was demonstrated by ELISA. CONCLUSION: The obtained gene engineering antibody against r-EgB may have potential implications in immunological treatment and drug targeting delivery.展开更多
[目的]探讨重组人白细胞介素-12（recombinant human interleukin-12,rhIL-12）在胰腺癌放射治疗放射防护中发挥的作用。[方法]回顾性分析52例胰腺癌患者,根据治疗方式分为单纯放疗组和rhIL-12＋放疗组。两组患者均采用射波刀治疗,放射...[目的]探讨重组人白细胞介素-12（recombinant human interleukin-12,rhIL-12）在胰腺癌放射治疗放射防护中发挥的作用。[方法]回顾性分析52例胰腺癌患者,根据治疗方式分为单纯放疗组和rhIL-12＋放疗组。两组患者均采用射波刀治疗,放射剂量30~35Gy,r IL-12＋放疗组放疗后予以150ng/kg rhIL-12干预治疗。两组分别于0h、12h、3d、7d、14d、21d、28d采用流式细胞术检测两组患者细胞免疫功能相关指标CD4/CD8比值、CD45、CD56数值变化;ELISA法检测血清细胞因子IFN-γ表达水平;分析评价两组患者外周血细胞计数变化、近期治疗疗效和胃肠道相关不良反应。[结果]rhIL-12＋放疗组患者免疫相关分子CD4/CD8比值、CD45和CD56表达12h内均存在一过性降低现象,随治疗时间的推移逐渐回升,于治疗第14d达到峰值且恢复,改善趋势明显优于单纯放疗组（P〈0.05）;细胞因子IFN-γ表达随时间变化逐渐递增,于14d达峰值,较单纯放疗组具有显著性差异（P〈0.05）。血液标本检测显示两组患者治疗前红细胞（RBC）、白细胞（WBC）、血小板（PLT）、中性粒细胞（NEUT）计数以及血红蛋白（Hb）含量均无显著性差异（P〉0.05）;在治疗14d时rhIL-12＋放疗组患者WBC、PLT和NEUT计数较单纯放疗组均具显著性差异（P〈0.05）,而RBC计数和Hb含量则差异无统计学意义（P〉0.05）;rhIL-12＋放疗组患者近期治疗疗效显著（P=0.046）;两组患者胃肠道不良反应主要表现为恶心呕吐和腹泻,rhIL-12＋放疗组发生率均低于单纯放疗组（P=0.001;P=0.012）。[结论]rhIL-12可有效地促进胰腺癌放射治疗患者造血功能和免疫机能的恢复和重建,并能降低放射治疗不良反应的发生率,提高治疗疗效。展开更多
文摘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.
文摘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.
基金The project was supported by a grant from the National Natural Science Fundation of China (No. 39860078) and Xinjiang Natural Science Fundation China (No. 200221101).
文摘OBJECTIVE: To identify a gene engineering antibody against cystic echinococcosis in liver. METHODS: A single chain of variable fragment of human antibodies (ScFvs) was selected from the library by using affinity selection technique with the recombinant antigen on solid surface. The positive clones were demonstrated by ELISA and their DNA sequences were also determined. RESULTS: The DNA sequence data showed that the antibody gene is composed of 768bp. In addition, a specific combination capacity with recombinant Echinococcus granulosus antigen B (r-EgB) was demonstrated by ELISA. CONCLUSION: The obtained gene engineering antibody against r-EgB may have potential implications in immunological treatment and drug targeting delivery.
文摘[目的]探讨重组人白细胞介素-12（recombinant human interleukin-12,rhIL-12）在胰腺癌放射治疗放射防护中发挥的作用。[方法]回顾性分析52例胰腺癌患者,根据治疗方式分为单纯放疗组和rhIL-12＋放疗组。两组患者均采用射波刀治疗,放射剂量30~35Gy,r IL-12＋放疗组放疗后予以150ng/kg rhIL-12干预治疗。两组分别于0h、12h、3d、7d、14d、21d、28d采用流式细胞术检测两组患者细胞免疫功能相关指标CD4/CD8比值、CD45、CD56数值变化;ELISA法检测血清细胞因子IFN-γ表达水平;分析评价两组患者外周血细胞计数变化、近期治疗疗效和胃肠道相关不良反应。[结果]rhIL-12＋放疗组患者免疫相关分子CD4/CD8比值、CD45和CD56表达12h内均存在一过性降低现象,随治疗时间的推移逐渐回升,于治疗第14d达到峰值且恢复,改善趋势明显优于单纯放疗组（P〈0.05）;细胞因子IFN-γ表达随时间变化逐渐递增,于14d达峰值,较单纯放疗组具有显著性差异（P〈0.05）。血液标本检测显示两组患者治疗前红细胞（RBC）、白细胞（WBC）、血小板（PLT）、中性粒细胞（NEUT）计数以及血红蛋白（Hb）含量均无显著性差异（P〉0.05）;在治疗14d时rhIL-12＋放疗组患者WBC、PLT和NEUT计数较单纯放疗组均具显著性差异（P〈0.05）,而RBC计数和Hb含量则差异无统计学意义（P〉0.05）;rhIL-12＋放疗组患者近期治疗疗效显著（P=0.046）;两组患者胃肠道不良反应主要表现为恶心呕吐和腹泻,rhIL-12＋放疗组发生率均低于单纯放疗组（P=0.001;P=0.012）。[结论]rhIL-12可有效地促进胰腺癌放射治疗患者造血功能和免疫机能的恢复和重建,并能降低放射治疗不良反应的发生率,提高治疗疗效。
