According to the traditional immunization procedure, after the first injection of the sample A (emulsion of aimed antigen and Freund's complete adjuvant) to immunize rabbit, successive injections of the sample B (...According to the traditional immunization procedure, after the first injection of the sample A (emulsion of aimed antigen and Freund's complete adjuvant) to immunize rabbit, successive injections of the sample B (emulsion of aimed antigen and Freund's incomplete adjuvant) were followed every 2-4 weeks. In general,high titer of the corresponding polyclonal antisera will be observed after 4-5 injections of sample B in 3-4months. This report presents a simply modified procedure that was able to stimulate the antisera formation in one month and achieve enough avidity to satisfy either Western blot or immunohistochemistry analysis.It just applied an additional injection of the sample A to the rabbit at the 3rd day after the primary immunization injection. You could gain the high titer of the antisera right after the first sample B injection in one month. This method has produced the desired results in three different recombinant antigens with different molecular weight (5.9 KD-55 KD) expressed from prokaryotic or eukaryotic cells.展开更多
BACKGROUND Chronic atrophic gastritis(CAG)is a common disease of the digestive system with pathological characteristics of a decreasing number,or disappearance,of inherent glands of the gastric mucosa.CAG has been def...BACKGROUND Chronic atrophic gastritis(CAG)is a common disease of the digestive system with pathological characteristics of a decreasing number,or disappearance,of inherent glands of the gastric mucosa.CAG has been defined as a precancerous condition of gastric cancer.Intestinal metaplasia or intraepithelial neoplasia accompanying atrophied glands of the stomach is regarded as one of the most important precancerous lesions of gastric cancer.As a common malignant tumour,gastric cancer remains without a satisfactory therapy and its pathogenesis remains unclear,seriously threatening human life.Therefore,some scholars have proposed to prevent the incidence of gastric cancer by avoiding precancerous lesions.If CAG can be reversed,the incidence of gastric cancer can be substantially reduced.To reverse and prevent CAG and study its pathogenesis and therapy,it is necessary to develop an ideal,safe,stable,animal model.AIM To study a rapid,stable,and safe method of establishing a mouse model of human CAG.METHODS Six-week-old Kunming mice were divided into a phosphate buffered solution control group,a Helicobacter pylori(H.pylori)group,an N-methyl-N'-nitroguanidine(MNNG)group,an ammonia water group,and a group combining H.pylori,MNNG,and ammonia water(hereinafter referred to as the combined group).The mice were administrated with drinking water containing ammonia or infected with H.pylori through gavage.At the 30th,60th,90th,and 120th day after the last H.pylori infection,mice were selected randomly to collect their gastric mucosa for hematoxylin eosin staining,terminal nick-end labelling staining detection,and immunohistochemical staining for Bax and Bcl-2.In addition,H.pylori was isolated,cultured,and identified,and its extent of colonisation calculated.Blood was collected to detect inflammatory factors interleukin(IL)-1β,IL-8,and tumor necrosis factor(TNF)-αand immune function markers CD4 and CD8 to confirm successful establishment of the CAG model.RESULTS The combined group showed slight CAG at the 90th day and moderat展开更多
基金We would like to thank Peifang Ping,BaozhenPeng and Dr.Xinxiu Yang for their helps in im-munization and ELISA.This work was funded bythe National Natural Sciences Foundation of China,No.39893320,the"973"Basic Research FundingScheme of China(G 199905590
文摘According to the traditional immunization procedure, after the first injection of the sample A (emulsion of aimed antigen and Freund's complete adjuvant) to immunize rabbit, successive injections of the sample B (emulsion of aimed antigen and Freund's incomplete adjuvant) were followed every 2-4 weeks. In general,high titer of the corresponding polyclonal antisera will be observed after 4-5 injections of sample B in 3-4months. This report presents a simply modified procedure that was able to stimulate the antisera formation in one month and achieve enough avidity to satisfy either Western blot or immunohistochemistry analysis.It just applied an additional injection of the sample A to the rabbit at the 3rd day after the primary immunization injection. You could gain the high titer of the antisera right after the first sample B injection in one month. This method has produced the desired results in three different recombinant antigens with different molecular weight (5.9 KD-55 KD) expressed from prokaryotic or eukaryotic cells.
基金Supported by National Natural Science Foundation of China,No.31460023Science Research and Technology Development Project of Guangxi,No.1598025-33
文摘BACKGROUND Chronic atrophic gastritis(CAG)is a common disease of the digestive system with pathological characteristics of a decreasing number,or disappearance,of inherent glands of the gastric mucosa.CAG has been defined as a precancerous condition of gastric cancer.Intestinal metaplasia or intraepithelial neoplasia accompanying atrophied glands of the stomach is regarded as one of the most important precancerous lesions of gastric cancer.As a common malignant tumour,gastric cancer remains without a satisfactory therapy and its pathogenesis remains unclear,seriously threatening human life.Therefore,some scholars have proposed to prevent the incidence of gastric cancer by avoiding precancerous lesions.If CAG can be reversed,the incidence of gastric cancer can be substantially reduced.To reverse and prevent CAG and study its pathogenesis and therapy,it is necessary to develop an ideal,safe,stable,animal model.AIM To study a rapid,stable,and safe method of establishing a mouse model of human CAG.METHODS Six-week-old Kunming mice were divided into a phosphate buffered solution control group,a Helicobacter pylori(H.pylori)group,an N-methyl-N'-nitroguanidine(MNNG)group,an ammonia water group,and a group combining H.pylori,MNNG,and ammonia water(hereinafter referred to as the combined group).The mice were administrated with drinking water containing ammonia or infected with H.pylori through gavage.At the 30th,60th,90th,and 120th day after the last H.pylori infection,mice were selected randomly to collect their gastric mucosa for hematoxylin eosin staining,terminal nick-end labelling staining detection,and immunohistochemical staining for Bax and Bcl-2.In addition,H.pylori was isolated,cultured,and identified,and its extent of colonisation calculated.Blood was collected to detect inflammatory factors interleukin(IL)-1β,IL-8,and tumor necrosis factor(TNF)-αand immune function markers CD4 and CD8 to confirm successful establishment of the CAG model.RESULTS The combined group showed slight CAG at the 90th day and moderat
