目的:中国角膜移植供体缺口巨大,可通过角膜成分移植来扩大供体角膜来源。本研究旨在通过观察采用不同储存介质保存的血清乙型肝炎表面抗原(hepatitis B surface antigen,HBsAg)阳性捐献者角膜中HBsAg的表达和分布,以评估板层角膜移植...目的:中国角膜移植供体缺口巨大,可通过角膜成分移植来扩大供体角膜来源。本研究旨在通过观察采用不同储存介质保存的血清乙型肝炎表面抗原(hepatitis B surface antigen,HBsAg)阳性捐献者角膜中HBsAg的表达和分布,以评估板层角膜移植的可行性和安全性。方法:本研究的研究对象为2019年12月至2021年3月间临床捐献的保存在中南大学湘雅三医院眼库的10枚角膜(来自6例捐献者)。采用酶联免疫吸附分析的方法检测血清HBsAg的表达,聚合酶链反应(polymerase chain reaction,PCR)法检测血清HBsAg阳性捐献者血中乙型肝炎病毒(hepatitis B virus,HBV)DNA滴度,免疫荧光染色检测角膜中的HBsAg的表达及其在角膜各层的定位,对血中HBV DNA滴度和角膜中HBsAg荧光强度进行相关性分析。结果:4枚来自2例血清HBsAg阳性捐献者的角膜、1枚来自1例血清HBsAg阴性捐献者的角膜均短期(<2周)保存在中期保存液中,2枚来自1例血清HBsAg阳性捐献者的角膜和2枚来自1例血清HBsAg阳性捐献者的角膜分别在甘油中保存1个月和1年,1枚来自1例血清HBsAg阴性捐献者的角膜在甘油中保存1年。免疫荧光染色结果显示:采用中期保存液短期(<2周)保存的2枚角膜上皮层和1枚角膜内皮层、甘油保存1年的1枚角膜上皮细胞均可见HBsAg阳性信号。免疫荧光染色半定量分析结果显示:角膜上皮层和内皮层中HBsAg荧光强度差异有统计学意义(P=0.012),不同捐献者及保存方法的角膜上皮中HBsAg荧光强度的差异无统计学意义(P>0.05)。血中HBV DNA滴度与角膜上皮中的HBsAg阳性表达呈正相关(R^(2)=0.707,P=0.036)。结论:HBsAg血清阳性捐献者的供体角膜存放于中期保存液或在甘油中长期保存后,上皮层或内皮层仍有HBsAg的阳性表达,但角膜基质层中均无HBsAg的阳性表达。血中HBV DNA滴度高表达的捐献者,其角膜中HBsAg表达阳性的可能性越高。本研究为进一步研究HBsAg阳性供体角膜用于板层角膜移植的可能性奠定了基础。展开更多
We have synthesized a novel formulation of nanoparticulate（NP） alum adjuvant specifically in the cationic water-in-oil microemulsions of water/benzalkonium bromide（BB）, octyl alcohol/cyclohexane at 30 ℃, which wa...We have synthesized a novel formulation of nanoparticulate（NP） alum adjuvant specifically in the cationic water-in-oil microemulsions of water/benzalkonium bromide（BB）, octyl alcohol/cyclohexane at 30 ℃, which was characterized to be crystalline Al（OH）3 with average particle diameter about 70 nm by the analysis of X-ray diffraction and to have a spherical shape structure by TEM photography analysis. Then adsorption degree was assayed. The OD492 of the NP is about 10 times higher than the bulk（BK） alum adjuvant. Compared to bulk alum adjuvant, we studied the immune effect of the NP alum adjuvant with HBsAg vaccine, which was injected intraperitoneally into Guinea pig, and serum antibody titers of the first and second week after immunization were higher in NP group than BK group by ELISA（P<0\^01, P<0\^05）; Groin immuno-injection intraperitoneally into Balb/c mice, was performed once or there times, prolife responses of spleen cells were determined by （）3H thymidine incorporation; non-specific phagocytosis of macrophage of abdominal cavity was measured by lack of color Malachite Green and IL-2 was assayed by activation mouse spleen cells. It is indicated that all cellular immunity bioassay results were higher in NP group than in BK group, except non-specific phagocytosis of macrophage of abdominal cavity by there times immunity. However, there was no significant differerces. Thus, nanoparticulate alum adjuvant can be employed as a more effective adjuvant for HBsAg vaccine than bulk alum adjuvant in the earlier period of post-immunity.展开更多
文摘We have synthesized a novel formulation of nanoparticulate（NP） alum adjuvant specifically in the cationic water-in-oil microemulsions of water/benzalkonium bromide（BB）, octyl alcohol/cyclohexane at 30 ℃, which was characterized to be crystalline Al（OH）3 with average particle diameter about 70 nm by the analysis of X-ray diffraction and to have a spherical shape structure by TEM photography analysis. Then adsorption degree was assayed. The OD492 of the NP is about 10 times higher than the bulk（BK） alum adjuvant. Compared to bulk alum adjuvant, we studied the immune effect of the NP alum adjuvant with HBsAg vaccine, which was injected intraperitoneally into Guinea pig, and serum antibody titers of the first and second week after immunization were higher in NP group than BK group by ELISA（P<0\^01, P<0\^05）; Groin immuno-injection intraperitoneally into Balb/c mice, was performed once or there times, prolife responses of spleen cells were determined by （）3H thymidine incorporation; non-specific phagocytosis of macrophage of abdominal cavity was measured by lack of color Malachite Green and IL-2 was assayed by activation mouse spleen cells. It is indicated that all cellular immunity bioassay results were higher in NP group than in BK group, except non-specific phagocytosis of macrophage of abdominal cavity by there times immunity. However, there was no significant differerces. Thus, nanoparticulate alum adjuvant can be employed as a more effective adjuvant for HBsAg vaccine than bulk alum adjuvant in the earlier period of post-immunity.
