Regulatory properties of brain glutamate decarboxylase (GAD): the apoenzyme of GAD is present principally as the smaller of two molecular forms of GAD in brain

The apoenzyme of glutamate decarboxylase [enzyme without bound cofactor, pyridoxal 5'-phosphate (pyridoxal-P)] serves as a reservoir of inactive glutamate decarboxylase (GAD) that can be activated when additional GABA synthesis is required. We have investigated which of two molecular forms of GAD is present as apoenzyme in synaptosomes and in cortex, caudate nucleus, hippocampus, and cerebellum of rat brain. Endogenous glutamate apodecarboxylase (apoGAD) was labeled by incubating extracts of synaptosomes or punches of each region with 32P-pyridoxal-P, followed by reduction with NaBH4, to link covalently the 32P-pyridoxal-P to GAD. Proteins were separated by SDS-PAGE. Punches from all four brain regions and forebrain synaptosomes contained two forms of GAD with apparent Mrs of 63 and 65 kDa as identified by immunoblotting with four antiGAD sera. Punches and synaptosomes contained a major 32P-pyridoxal-P-labeled band with an apparent Mr of 63 kDa that was stained on immunoblots by the antiGAD serum 1440 and the monoclonal antibody GAD-6, and a minor labeled band at 65 kDa that was stained by the 1440, 6799, and K2 antisera. Synaptosomes contained remarkably few other strongly labeled proteins, but punches contained several other labeled bands. Three additional lines of evidence indicate that the labeled 63-kDa protein is apoGAD: (1) it was purified by immunoaffinity chromatography with the GAD-1 monoclonal antibody; (2) it yielded one major labeled peptide when digested with chymotrypsin, and that peptide appeared identical in peptide-mapping experiments to the labeled active-site peptide isolated from chromatographically prepared rat brain GAD; and (3) its labeling was selectively blocked by 4-deoxypyridoxine 5'-phosphate, a competitive inhibitor of the binding of pyridoxal-P to GAD.(ABSTRACT TRUNCATED AT 250 WORDS)     

来源于骨髓和脐带血的基质细胞基本特性的比较

目的比较骨髓与脐带血细胞体外培养基质细胞的基本特性,为基质细胞的选择应用提供依据.方法用Dexter长期培养体系培养骨髓和脐带血基质细胞,以细胞增殖、细胞形态、细胞化学染色、细胞表面及基质细胞支持另一骨髓细胞形成的鹅卵石造血区(CAFC),长期培养起始细胞(LTC-IC)为指标,比较两者的生长特性、成分及功能.结果①细胞生长特性:出现贴壁细胞时间,骨髓细胞为培养3d,脐带血细胞为培养5～6d;细胞融合成片时间,骨髓细胞为培养10～14d,脐带血细胞为培养12～18d;第21天细胞增殖数,骨髓比脐带血细胞增殖少;②细胞成分:21d培养后细胞成分,骨髓来源者以成纤维细胞为主,其次是巨噬细胞与内皮细胞,脂肪细胞最少;脐带血细胞来源者,以巨噬细胞为主,其次是内皮细胞、成纤维细胞,偶见脂肪细胞;细胞化学与上述结果基本一致;细胞表面抗原检测,CD14、CD45的表达骨髓细胞明显低于脐带血细胞;③细胞功能:骨髓来源的基质细胞较脐带血细胞的基质细胞支持另一骨髓细胞形成的CAFC和LTC-IC明显多.结论①生长特征:形成贴壁细胞时间骨髓较脐带血短,骨髓细胞比脐带血细胞有核细胞数增殖快、持续时间相对短;②细胞成分特性:骨髓来源形成的基质细胞以成纤维细胞为主,脐带血来源者以巨噬细胞为主;③细胞功能特性:骨髓细胞形成的贴壁细胞较脐带血细胞形成的贴壁细胞更利于CAFC、LTC-IC生长.     

Efficacy of radiology of the esophagus for evaluation of dysphagia

The efficacy of radiology in evaluating dysphagia was studied in 86 patients by comparison to endoscopic findings. In the 66 patients with endoscopic abnormalities radiology was correct in 54, for a sensitivity of 82%. Sensitivity of radiology improved to 95% if mild esophagitis was excluded. In the 20 patients with normal endoscopy, radiology was normal in 18 (90%). Thus radiology proved to be a reliable means of evaluating the esophagus in patients with dysphagia.     

Functional recognition of the neuronal tyrosine hydroxylase cAMP regulatory element in different cell types.

Transient transfection of pLB2CAT constructs bearing short synthetic oligonucleotides derived either from the tyrosine hydroxylase (TH) promoter or other sources was used to examine functional cAMP regulatory element (CRE) activity in a variety of cell lines. The region containing only the putative TH CRE was found to be as or more effective in conferring cAMP responsiveness onto pLB2CAT (which employs the TK promoter) than the immediate 272 bp region of the TH promoter. Increases in CAT activity of 10- to 20-fold were observed in JEG-3 cells with a single insert of the TH CRE region (-31 to -54) in pLB2CAT, and the presence of a second insert generated only a modest further increase. This construct also responded to cAMP in 4 other cell lines tested but the degree of increase was less dramatic. Inserts containing the consensus 8 bp CRE motif embedded in other natural or artificial contexts served generally as weak functional CREs in all cell lines tested. In vitro analysis revealed that a specific protein-DNA complex apparently containing a single protein with a MW of 45-50 kDa was formed equally well with JEG-3 cell nuclear extract and CRE-bearing-TH and other fragments which produced dramatically different cAMP effects in vivo. These results suggest specificity in the effects of cAMP on different CREs which are dictated by contextual differences.     

选择性环氧化酶—2抑制剂的研究现状及应用评价

选择性环氧化酶（COX）-2抑制剂可有效地治疗炎症，同时避免或减轻由于抑制COX-1而导致的毒副作用。有乐观的临床应用前景，但其不良反应亦不容忽视，寻找临床评价好的选择性COX-2抑制剂是当前非甾体类抗类药物研究的重点。     

烯烃的热力学性质的拓扑研究

目的：寻找一种性能优良的拓扑指数，方法：根据分子图的邻接矩阵和距离矩阵提出一种新的拓扑指数P1^*,P2^*,P3^*,p^4*。结果，拓扑指数P1^*,P2^*,P3^*,p^4*与烯烃的热力学函数－△Hf(g),S(g),△Gf(g)均有相关性，其复相关系数均在99．6％以上。结论：拓扑指数P1^*,P2^*,P3^*,p^4*与烯烃的热力学函的多种物理化学性质均有良好的相关性。     

保留肾单位手术治疗早期小肾癌21例临床分析

OBJECTIVE: To evaluate the clinical effects of nephron-sparing surgery in patients with early-stage small renal cell carcinoma. METHODS: Nephron-sparing surgery was performed in 21 patients with renal cell carcinoma including 1 with solitary kidney, 3 with unilateral tumor and contralateral renal compromise, and 17 with unilateral tumor and normal contralateral kidney. The diameter of the tumors ranged from 1.5 to 6.0 cm, with a mean of 2.8 cm. The tumor diameter in 17 patients with normal contralateral kidney was less than 4 cm (mean 2.5 cm) and the average diameter in 4 patients with contralateral renal compromise was 4.2 cm. Sixteen cases were in stage T(1), 4 in stage T(2), and 1 in stage T(3). Of the 21 patients, 4 underwent tumor enucleation, 10 polar nephrectomy and 7 wedge resection. RESULTS: All patients were followed up for an average of 40.8 months (7 to 66 months). One patient suffered a right lung and mediastinum metastasis 3 years after the surgery later and 1 with chronic glomerulonephritis required dialysis 27 months after the operation. No surgical complication or local recurrence were found in other patients. CONCLUSION: As a safe and effective therapy for early-stage small renal cell carcinoma, nephron-sparing surgery can be considered as the gold-standard therapy for patients with lesions less than 4 cm in T(1) and T(2) stages of localized unilateral tumor with normal contralateral kidney.     

外伤性延迟性脾破裂的诊断和治疗（附21例报告）

目的探讨外伤性延迟性脾破裂的发病规律、临床特点、诊断和治疗方法。方法结合国内外资料及本组病例进行回顾性分析。结果明确诊断16例，误诊为肝破裂2例，宫外孕破裂2例，脾肿瘤1例。21例均手术治疗，行脾切除14例，其中保留副脾2例；脾切除加自体脾组织网膜内移植术3例；脾缝合修补术3例；脾部分切除术1例。死亡1例，原因有就诊晚、失血性休克。结论本病由于腹腔内出血与受伤时间间隔长，容易误诊。诊断除依靠病史、临床表现外，应及时进行腹腔穿刺、B超及CT检查。治疗以脾切除为主，可根据病情、脾破裂的程度以及是否有合并伤等情况采取保脾手术。