目的探讨制备脂质体包裹重组幽门螺杆菌过氧化氢酶(Kat)口服疫苗的方法,并用Hp感染的小鼠模型评价其在预防Hp感染中的作用。方法将PET-22(+)/Kat在BL21(DE3)大肠杆菌表达,表达Kat的包涵体经洗涤、变性、复性,采用Q Sepharose Fast Flow...目的探讨制备脂质体包裹重组幽门螺杆菌过氧化氢酶(Kat)口服疫苗的方法,并用Hp感染的小鼠模型评价其在预防Hp感染中的作用。方法将PET-22(+)/Kat在BL21(DE3)大肠杆菌表达,表达Kat的包涵体经洗涤、变性、复性,采用Q Sepharose Fast Flow阴离子交换层析和Sephacryl S-200凝胶过滤层析分离纯化Kat重组蛋白,用薄膜分散法制备以卵磷脂和胆固醇为膜组分包裹的Kat重组蛋白口服疫苗,并用透射电镜测定其粒径。BALB/c小鼠分为5组,分别通过灌胃方法给予PBS、空白脂质体、Kat重组蛋白+CT、脂质体包裹Kat重组蛋白、脂质体包裹Kat重组蛋白和CT,每周1次共4次,末次攻击2周再用活Hp攻击3次,3周后处死小鼠,行胃组织快速尿素酶试验、Hp的定植半定量、炎症程度及其炎症活动度的评分。结果以包涵体为主的表达产物占全菌总蛋白的24.4%,经纯化获得纯度为95%的重组蛋白,制备的脂质体粒径为0.7±0.4μm。PBS组和空白脂质体组保护率均为0,而Kat重组蛋白+CT组、脂质体包裹Kat重组蛋白组、脂质体包裹Kat重组蛋白和CT组的保护率分别为73.3%、66.7%和86.7%,且均能使免疫小鼠胃粘膜Hp感染数目明显减少,炎症反应减轻。结论口服脂质体部能分代替免疫佐剂的作用,将其作为Hp疫苗的免疫佐剂,具有广泛的应用前景。展开更多
Our purpose was to investigate associations between adenosine deaminase (ADA), superoxide dismutase (SOD), and catalase (CAT) activities and H. pylori. Ninety-nine patients were studied. Eight antral mucosal biopsies ...Our purpose was to investigate associations between adenosine deaminase (ADA), superoxide dismutase (SOD), and catalase (CAT) activities and H. pylori. Ninety-nine patients were studied. Eight antral mucosal biopsies were taken for biochemical assessment of ADA, CAT, and SOD activity and histological assessment. H. pylori density was evaluated according to the updated Sydney system. Patients were divided into three groups according to Sydney classification. ADA activity was found to be higher in patients having H. pylori in the present study. Also, ADA activity was higher in patients with a severe density of H. pylori. SOD level was found to be significantly higher with increased H. pylori density in our study (P < 0.05). In addition, SOD activity was higher in H. Pylori-positive than H. pylori-negative patients. We did not find CAT activity in some antral tissue specimens. The significantly high levels of ADA activity in patients with H. pylori infectionmay reflect the regulator role of ADA in acid secretion. The higher ADA level with increased H. pylori density and H. pylori positivity indicate the probable malign lymphoid process of the stomach. But these findings must be confirmed with larger studies that include different gastric lesions.展开更多
文摘目的探讨制备脂质体包裹重组幽门螺杆菌过氧化氢酶(Kat)口服疫苗的方法,并用Hp感染的小鼠模型评价其在预防Hp感染中的作用。方法将PET-22(+)/Kat在BL21(DE3)大肠杆菌表达,表达Kat的包涵体经洗涤、变性、复性,采用Q Sepharose Fast Flow阴离子交换层析和Sephacryl S-200凝胶过滤层析分离纯化Kat重组蛋白,用薄膜分散法制备以卵磷脂和胆固醇为膜组分包裹的Kat重组蛋白口服疫苗,并用透射电镜测定其粒径。BALB/c小鼠分为5组,分别通过灌胃方法给予PBS、空白脂质体、Kat重组蛋白+CT、脂质体包裹Kat重组蛋白、脂质体包裹Kat重组蛋白和CT,每周1次共4次,末次攻击2周再用活Hp攻击3次,3周后处死小鼠,行胃组织快速尿素酶试验、Hp的定植半定量、炎症程度及其炎症活动度的评分。结果以包涵体为主的表达产物占全菌总蛋白的24.4%,经纯化获得纯度为95%的重组蛋白,制备的脂质体粒径为0.7±0.4μm。PBS组和空白脂质体组保护率均为0,而Kat重组蛋白+CT组、脂质体包裹Kat重组蛋白组、脂质体包裹Kat重组蛋白和CT组的保护率分别为73.3%、66.7%和86.7%,且均能使免疫小鼠胃粘膜Hp感染数目明显减少,炎症反应减轻。结论口服脂质体部能分代替免疫佐剂的作用,将其作为Hp疫苗的免疫佐剂,具有广泛的应用前景。
文摘Our purpose was to investigate associations between adenosine deaminase (ADA), superoxide dismutase (SOD), and catalase (CAT) activities and H. pylori. Ninety-nine patients were studied. Eight antral mucosal biopsies were taken for biochemical assessment of ADA, CAT, and SOD activity and histological assessment. H. pylori density was evaluated according to the updated Sydney system. Patients were divided into three groups according to Sydney classification. ADA activity was found to be higher in patients having H. pylori in the present study. Also, ADA activity was higher in patients with a severe density of H. pylori. SOD level was found to be significantly higher with increased H. pylori density in our study (P < 0.05). In addition, SOD activity was higher in H. Pylori-positive than H. pylori-negative patients. We did not find CAT activity in some antral tissue specimens. The significantly high levels of ADA activity in patients with H. pylori infectionmay reflect the regulator role of ADA in acid secretion. The higher ADA level with increased H. pylori density and H. pylori positivity indicate the probable malign lymphoid process of the stomach. But these findings must be confirmed with larger studies that include different gastric lesions.
