Effect of the expression of DRαEβNOD molecule on the development of insulitis and diabetes in the non-obese diabetic (NOD) mouse

Previous studies have shown that a transgenic I-Eα gene, the mouse homologue of human DRα gene, prevents the development of insulitis and hence of diabetes in NOD mice. To investigate the mechanism of this prevention, we generated two strains of NOD mice expressing DRαEβ molecule: DRα-24-NOD expressing DRαEβ molecule on thymic epithelial cells (TEC) and bone marrow-derived cells (BDC), and DRα-30-NOD expressing DRαEβ molecule only on the TEC, and these mice were monitored for disease development. Because the DRαEβ molecule reconstituted I-E controlled immune regulation, it would become clear which cell type, TEC or BDC, was responsible for the I-E-mediated disease protection. To our surprise, however, DRα-24-NOD developed insulitis and diabetes comparably to non-transgenic littermates. This suggested that the difference in structure between DRα and Eα molecules contributed to the difference in preventive effect on the development of insulitis and diabetes between DRα-24-NOD and Eα-NOD. In an analysis of the T cell proliferative responses to glutamic acid decarboxylase (GAD) 65-derived peptides which were known to be diabetogenic autoantigens, it was shown that DRα-24-NOD and NOD acquired comparable level of T cell response to GAD 509–528 but 5–10-fold higher response was observed in Eα-NOD. This suggested that I-A^NOD and EαEβ^NOD molecules could present GAD 509–528 peptide to T cells, while DRαEβ^NOD could not. Furthermore, T cells from DRα transgenic mice showed proliferative response to antigen-presenting cells from Eα transgenic mice in primary mixed lymphocyte reaction. This also suggested that the EαEβ molecule does differ in structure and peptide binding from the DRαEβ molecule. Present data suggested a possibility that the T cell repertoire selection, or the T cell response to GAD 65 and/or other unknown antigens specifically mediated by I-E molecule, may contribute to the prevention of disease development in Eα-NOD.     

Prostaglandins and cyclic AMP in epidermis

Prostaglandins E1 and E2 stimulate cyclic AMP accumulation in pig epidermis and in human epidermis from patients with psoriasis. Prostaglandins A1,A2 and F2alpha are relatively ineffective. The fact that this stimulation is not inhibited by a beta-blocker (propranolol) and that the stimulation by prostaglandin E2 and adrenaline is additive indicates that each drug acts independently on the epidermal adenyl cyclase system. In other words, prostaglandins E1 and E2 act on a site other than the beta-receptor of adenyl cyclase in epidermis. The stimulation by prostaglandins E1 and E2 is not additive; hence they probably act on the same site. Concentrations of prostaglandin E above 3X10(-7) M are effective in causing stimulation. This concentration may be within the physilogical range and the contribution of endogenous prostaglandin levels in the control of intracellular cyclic AMP levels cannot be disregarded.     

仙人掌中一个新α-吡喃酮成分的分离与结构鉴定

目的　研究仙人掌的化学成分。方法　应用多种柱色谱方法进行分离和纯化 ,NMR和MS等波谱解析化学结构。结果　从仙人掌肉质茎的乙醇提取物中分离出 6个化合物 ,其结构分别鉴定为 :3 O 甲基异鼠李黄素 (1)、4 乙氧基 6 羟甲基 α 吡喃酮 (2 )、正十七醇 (3)、香草酸 (4)、异鼠李黄素 3 O 鼠李糖苷 (5 )和芦丁 (6 )。结论　化合物2为新化合物 ,化合物 1、3、4、5均为首次从本属植物中分离得到 ,化合物 6为本植物首次分离得到。     

Effects of cytokines on hematopoietic progenitor cells in cord blood,in bone marrow,and in peripheral blood mobilized by chemotherapy and G-CSF

We compared the effects of various combinations of cytokines (stem cell factor [SCF], interleukin [IL] ?3, IL-6, granulocyte-colony stimulating factor [G-CSF], erythropoietin [EPO]) among the growth of human hematopoietic progenitor cells from cord blood (CB), bone marrow (BM), and peripheral blood mononuclear cells (MNC) mobilized by chemotherapy and G-CSF (PB) in a semi-solid medium. Macroscopic colonies, that were visible to the naked eye, were formed from PB-MNC within 1 week even without cytokines. They consisted of blasts containing macrophage-like cells with immature nuclei on Wright stain, and were strongly accelerated by IL-3. Macroscopic colonies were also formed from CB-MNC. However, they appeared after 1–3 weeks and synergistic effects of SCF with other cytokines, especially EPO, were prominent. Macroscopic colonies were not formed from BM-MNC. Granulocyte-colony stimulating factor was effective in increasing colony forming units of granulocyte macrophage from BM-MNC and they appeared between 1 and 2 weeks. These results suggested that the quality of hematopoietic progenitor cells was different among blood sources. This might lead to different bone marrow recovery patterns after transplantation of each blood source. The appropriate cytokines should be added to evaluate their exact potential.     

Ex vivo expansion of umbilical cord blood hematopoietic progenitor cells by combinations of cytokines

Ex vivo expansion of hematopoietic progenitor cells in the umbilical cord blood mononuclear cells (CB-MNC) was investigated in liquid culture system with various combinations of cytokines (stem cell factor [SCF], interleukin [IL]-3, IL-6, granulocyte-colony stimulating factor [G-CSF], erythropoietin [EPO], and interferon [INF]-γ). Non-lineage-committed hematopoietic progenitor cells and lineage committed hematopoietic progenitor cells were represented as CD34⁺CD38^? and CD34⁺CD38⁺ subpopulations, respectively. Although absolute CD34⁺CD38^? cell numbers decreased even in the presence of multicytokines, the combinations of SCF plus IL-6 and SCF plus IL-3 plus IL-6 plus INF-γ were significantly effective in maintaining CD34⁺CD38^? cells than the other combinations (P < 0.05). After 4 weeks of culture, CD34⁺CD38^? cells disappeared in all combinations of cytokines. Absolute CD34⁺CD38⁺ cell numbers increased in the presence of cytokines. Maximal expansion of CD34⁺CD38⁺ cells were observed in the combinations of SCF plus IL-3 plus IL-6 plus EPO (19.8 ± 3.3 -fold) and SCF plus IL-3 plus IL-6 plus G-CSF (18.3 ± 2.6). The combination of SCF plus IL-3 plus IL-6 was also effective to expand CD34⁺CD38⁺ cells (15.8 ± 3.9). However, the expansion was transient and they decreased to zero within 3 weeks. In the combinations of SCF plus IL-6 and SCF plus IL-3 plus IL-6 plus INF-γ, maximal expansion was inferior to the others but CD34⁺CD38⁺ cells were maintained more than 4 weeks. These results suggested that the indication of CBT can be expanded into older children by ex vivo augmentation of CB hematopoietic progenitor cells using multi-cytokines.     

Glomerular morphometry I: nephrotic syndrome in childhood

Glomerular morphometry was performed on needle biopsy specimens from 53 children with the nephrotic syndrome with minimal change (MC), focal global glomerulosclerosis (FGS) and segmental glomerulosclerosis (SGS). There were significantly fewer epithelial cells and more mesangial cells in SGS compared with MC and FGS. The number of epithelial cells decreased with age in MC and FGS, but was constantly low at all ages in SGS. There were no significant differences in differential cellularity between MC, with or without focal glomerular obsolescence or focal tubular atrophy, and FGS. Glomerular diameter increased with age in MC and FGS, but not in SGS. These findings support the view that SGS is a distinct entity rather than a variant of MC.     

Evaluation of Chemotherapy for Locally Unresectable Gastric Cancer

The effect of chemotherapy on locally unresectable gastric cancerwas evaluated on the basis of survival. The result showed asignificant effect of chemotherapy in the six-month survivalrate. Chemotherapy prolonged survival by at most two months.The result favored three major regimens, i.e., the FAMT (5-fluorouracil,cyclophosphamide, mitomycin C and chromomycin A³) combination,l-(2-tetrahydrofuryl)-5-fluorouracil (Ftorafur) alone and intra-aorticinfusion of 5-FU, as compared with the controls. However, survivalwas not very different among these regimens. In respect to factorscontrolling the background of gastric cancer, the effect ofchemotherapy was high in the cases in which the S (serosal involvement)and N (lymph node involvement) factors were dominant withoutthe association of the P (peritoneal dissemination) or H (hepaticmetastasis) factor. The participation of H factor resulted inthe worst prognosis, the patients receiving little help fromchemotherapy. Patients with P factor occasionally survived longer,especially with subsequent chemotherapy.     

多叶姜黄(Curcuma comosa Roxb.)的化学成分

目的对泰国产姜科姜黄属植物多叶姜黄(Curcuma comosaRoxb.)干燥根茎甲醇提取物的化学成分进行分离和结构鉴定。方法C.comosaRoxb.的甲醇提取物经乙酸乙酯-正丁醇-水依次萃取后,对各萃取部分采用正-反相硅胶柱色谱和制备型HPLC进行分离,经理化常数测定、核磁共振技术分析等方法鉴定了化合物的结构。结果分离得到8个化合物,分别被鉴定为异蓬莪术环二烯酮(isofuranodienone,1),蓬莪术环二烯酮(furanodienone,2),1(10)Z,4Z-蓬莪术二烯-6-酮[1(10)Z,4Z-furanodiene-6-one,3],泽泻醇(alismol,4),2,2,6-三甲基-1-氧螺[2,5]辛-5-烯-4-醇(2,2,6-trimethyl-1-oxaspiro[2,5]oct-5-en-4-ol,5),1-羟基-α,α,4-三甲基-3-环己烯-1-甲醇(1-hydroxy-α,α,4-trimethyl-3-cyclohexene-1-methanol,6),6-羟基-3(1-羟基-1-甲基乙基)-6-甲基-2-环己烯-1-酮(6-hydroxy-3(1-hydroxy-1-methylethyl)-6-methyl-2-cyclo-hexen-1-one,7),(1S,2S,4R)-1,8桉叶素-2-O-β-D-葡萄糖苷((1S,2S,4R)-2-hydroxy-1,8-cineoleβ-D-glucopyranoside,8)。结论化合物3~8为首次从该属植物中分离得到。     

仙人掌的化学成分研究

目的:研究民间药用植物仙人掌的化学成分。方法:应用多种柱色谱方法进行分离和纯化,NMR和MS等波谱解析化学结构。结果:从仙人掌肉质茎的乙醇提取物中分离出6个化合物,其结构鉴定为:仙人掌醇(1)、胡萝卜苷(2)、对羟基苯甲酸(3)、L-(-)-苹果酸(4)、仙人掌苷(5)和阿魏酸(6)。结论:化合物2,3,4,6均为首次从本属植物中分离得到,化合物1为本植物首次分离得到,化合物5为新化合物。