AN ATTEMPT TO INTRODUCE GLYCOBIOLOGICAL OR BIOCHEMICAL TECHNIQUES INTO FORENSICPATHOLOGICAL DIAGNOSIS

将糖生物学或生物化学技术引入法医病理诊断的尝试

• 批准号: 11670413
• 负责人: NISHI Katsuji
• 金额: $ 2.3万
• 依托单位: SHIGA UNIVERSITY OF MEDICAL SCIENCE
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11670413/
• 关键词: glycoconjugate; carbohydrate chain; degenerative deposition; histochemistry; brain disorder; レクチン; ドーパミン; ペプチダーゼ; 糖鎖構造物質

Lectin histochemical technique clearly revealed the existence of senile plaque, neurofibrillary tangles, corpora amylacea, and three types of carbohydrate depositions, that is, spherical, vascular and stratiform type, in the human brain from patients with brain disorder. The spherical type deposits were detected in the molecular layer of the dentate gyrus of the hippocampal formation of patients with schizophrenia. The vascular and stratiform type which showed amorphous in the hematoxylin eosin staining were detected in the white matter region in the cerebrum and cerebellum of patients with brain disorder, such as dementia, Down's syndrome or brain edema. Dipeptidyl peptidase III (DPIII) and alanyl aminopeptidase (AP) were purified from human liver cytosol, and antibodies against these enzymes were produced. Antibody against AP histochemically reacted with neurons and glial cells, and could stain the above mentioned amorphous depoditions. The depositions newly detected in this project may be results from the disturbance of glycogenesis or blood brain barrier in the brain and relate to brain disorder. Antibodies against serotonin 2A receptor and tyrosine hydroxylase revealed that many neurons in the human ventral tegment area and substantia nigra are doubly labeled and suggested that activity of human midbrain dopaminergic neurons might be strongly regulated via serotonin 2A receptors at the level of their originating nuclei.

凝集素组织化学技术清楚地显示了脑病患者脑内存在老年斑、神经原纤维缠结、淀粉体和三种类型的碳水化合物沉积，即球形、血管型和层状型。精神分裂症患者海马齿状回分子层可见球形沉淀物。在痴呆、唐氏综合征、脑水肿等脑功能障碍患者的大脑和小脑白质区，可见苏木精-伊红染色呈无定形的血管型和层状型。从人肝细胞胞浆中提纯了二肽基肽酶III(DPIII)和丙氨酰氨基肽酶(AP)，并制备了相应的抗体。抗AP抗体可与神经元和神经胶质细胞发生组织化学反应，并可对上述无定形细胞进行染色。本项目新检测到的沉积物可能是脑内糖生成障碍或血脑屏障障碍的结果，与脑功能障碍有关。抗5-羟色胺2A受体和酪氨酸羟基酶抗体显示，人中脑腹侧被盖区和黑质的许多神经元被双重标记，提示人中脑多巴胺能神经元的活动可能通过5-羟色胺2A受体在其起始核团水平上受到强烈的调节。

项目成果

Ohkubo I.,Li Y-H.,Maeda T.,Yamamoto Y.,Yamane T.,Nishi K.,: "Dipeptidyl peptidase III from rat liver cytosol : Purification, molecular cloningand immunohistochemical localization."Biol.Chem.. 380. 1421-1430 (1999)

Ohkubo I.、Li Y-H.、Maeda T.、Yamamoto Y.、Yamane T.、Nishi K.，：“来自大鼠肝细胞质的二肽基肽酶 III：纯化、分子克隆和免疫组织化学定位。”Biol.Chem.. 380. 1421

Y.Yamamoto,Y-H. Li,I.Ushiyama,A.Nishimura,I.Ohkubo,K.Nishi.: "Puromycin-sensitive alanyl aminopeptidase from human liver cytosol : purificationand characterization."Forensic Science Interational. 113. 143-146 (2000)

Y.山本，Y-H。

澤田園、西村明儒、牛山郁子、中川季子、山本好男、西克治: "鍍銀法と二種の免疫染色による三重染色の法医神経病理診断学への応用"法医病理. 6巻(印刷中). (2000)

Sono Sawada、Akihiro Nishimura、Ikuko Ushiyama、Kiko Nakakawa、Yoshio Yamamoto、Katsuharu Nishi：“使用镀银法的三重染色和两种类型的免疫染色在法医神经病理学诊断中的应用”《法医病理学》第 6 卷（印刷版）（2000 年）。

K.Ikemoto,R.Amano,A.Ishibe,A.Nishimura,K.Nishi,I.Nagatsu: "Quantitative analysis of tyrosine hydroxylase-, aromatic L-amino acid decarboxylase- or phenylethanoamine-N-methyltransferase-immnoreactive neurons in the human medullary C1 region."Acta Histochem

K.Ikemoto、R.Amano、A.Ishibe、A.Nishimura、K.Nishi、I.Nagatsu：“酪氨酸羟化酶、芳香族 L-氨基酸脱羧酶或苯乙醇胺-N-甲基转移酶免疫反应性神经元的定量分析

Y.Yamamoto, Y-H.Li, I.Ushiyama, A.Nishimura, I.Ohkubo, K.Nishi.: "Puromycin-sensitive alanyl aminopeptidase from human liver cytosol : purificationand characterization."Forensic Science Interational. 113. 143-146 (2000)

Y.Yamamoto、Y-H.Li、I.Ushiyama、A.Nishimura、I.Ohkubo、K.Nishi.：“人肝细胞质中的嘌呤霉素敏感丙氨肽酶：纯化和表征。”国际法医科学。