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管氏肿腿蜂毒液丝氨酸蛋白酶同源物SgSPH对寄主血淋巴酚氧化酶活性的影响 被引量:2
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作者 李丽芳 吴朝妍 +2 位作者 韩开健 吴国星 朱家颖 《昆虫学报》 CAS CSCD 北大核心 2021年第2期170-177,共8页
【目的】本研究旨在通过克隆表达管氏肿腿蜂Scleroderma guani毒液丝氨酸蛋白酶同源物(serine protease homologue,SPH)基因SgSPH,探索其编码的毒液蛋白对寄主血淋巴酚氧化酶活性的影响。【方法】利用RT-PCR技术克隆管氏肿腿蜂毒液SgSP... 【目的】本研究旨在通过克隆表达管氏肿腿蜂Scleroderma guani毒液丝氨酸蛋白酶同源物(serine protease homologue,SPH)基因SgSPH,探索其编码的毒液蛋白对寄主血淋巴酚氧化酶活性的影响。【方法】利用RT-PCR技术克隆管氏肿腿蜂毒液SgSPH基因的开放阅读框(ORF),采用生物信息学软件分析其基因序列结构特征;采用qPCR技术检测该基因在不同发育阶段(卵、低龄幼虫、高龄幼虫、老熟幼虫、吐丝幼虫、黄茧蛹、黑茧蛹和羽化后1-5 d成虫)和雌成虫不同组织(头部、胸部、去除毒液器官的腹部和毒液器官)中的相对表达量;利用载体pSUMO-Mut对该基因进行原核表达,用镍亲和层析柱纯化表达的重组蛋白,采用SDS-PAGE和Western blot对获得的重组蛋白进行鉴定;用酶活性测定方法,分析SgSPH重组蛋白对寄主黄粉虫Tenebrio molitor蛹血淋巴酚氧化酶活性的抑制作用。【结果】克隆获得管氏肿腿蜂毒液SgSPH基因(GenBank登录号:MT920663)的ORF,长798 bp,编码265个氨基酸,其中第1-20位氨基酸为信号肽,理论分子量为30.53 kD,等电点为9.59。多序列比对分析表明,SgSPH与其他寄生蜂毒液丝氨酸蛋白酶和SPH具有较低的氨基酸序列一致性(9%~17%),且缺乏保守的催化三联体。qPCR分析表明,SgSPH基因在管氏肿腿蜂成虫阶段和毒液器官中高表达。SDS-PAGE电泳和Western-blot检测发现,成功表达SgSPH重组蛋白,并纯化得到了高纯度SgSPH重组蛋白。酶活性检测结果显示,SgSPH能抑制寄主黄粉虫蛹血淋巴的酚氧化酶活性。【结论】结果提示管氏肿腿蜂毒液SgSPH具有干扰寄主酚氧化酶级联反应的功能。 展开更多
关键词 管氏肿腿蜂 毒液 丝氨酸蛋白酶同源物 原核表达 表达特征 酚氧化酶
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Nailfold Capillaroscopy Findings in Diabetic Patients (A Pilot Cross-Sectional Study)
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作者 Alireza Rajaei Pooneh Dehghan Zahra Farahani 《Open Journal of Pathology》 2015年第2期65-72,共8页
Background: Microcirculation is affected in diabetes mellitus and Microvascular abnormalities cause persistent diabetic complications. The aim of this study was nailfold capillaroscopic assessment to describe the path... Background: Microcirculation is affected in diabetes mellitus and Microvascular abnormalities cause persistent diabetic complications. The aim of this study was nailfold capillaroscopic assessment to describe the pathological changes (morphological and structural) in capillary of a large series of patients with type 1 and 2 diabetes. Methods: A cross-sectional study was carried out in a Nailfold Capillaroscopy Center (Tehran-Iran) between 2011 and 2014. The study included 235 types 1, 2 diabetic patients. All patients underwent 10 nailfolds capillaroscopy examinations. Microvascular architecture, disturbances capillary distribution, capillary morphology, capillary density, efferent/afferent limb ratio, subpapillary venular plexus, and morphological abnormalities were evaluated. Conclusions were stated as normalor scleroderma pattern. Results of patients’ capillaroscopic images were recorded and analyzed quantitatively and qualitatively. P value < 0.05 was considered as statistical significance. Results: of all participants with mean age 59.91 ± 12.39, 183 cases (77.9%) were female and 52 (22.1%) were male. Tortuosity of capillaries was more often observed in our subjects (235 cases) followed by angiogenesis (171 cases). Normal and early scleroderma patterns were observed in 195 (83.0%) and 40 cases (17.0%). Based on P values, altered micro vascular architecture, capillary distribution and capillary morphology were more frequent in patients with scleroderma pattern in comparison to patients with normal pattern (P value < 0.05). Morphological abnormalities except from neo formation capillary and mega capillary were also significantly more common in patients with scleroderma pattern than patients in counterpart group (P value < 0.05). Conclusion: Nailfold capillaroscopy as a non-invasive, diagnostic and prognostic method may potentially affect on diabetes outcome and control. 展开更多
关键词 Diabetes NAIL FOLD CAPILLAROSCOPY Normal pattern scleroderma patternDiabetes scleroderma pattern
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