Spectrophotometric assay for ornithine decarboxylase

A rapid and sensitive spectrophotometric assay for ornithine decarboxylase is described. It is based on the observation that the product of ornithine decarboxylase, putrescine, reacts with 2,4,6-trinitrobenzenesulfonic acid to give a colored product soluble in 1-pentanol whereas ornithine does not. The amount of putrescine produced by the enzyme was determined by measuring the absorbance of the 1-pentanol extract of the reaction mixture at 420 nm, and by comparing the results to those obtained by the trapping of 14CO2 and by HPLC assays. The three assays were found to be equivalent in sensitivity, with the spectrophotometric assay having the advantages of being relatively rapid, requiring only common laboratory equipment, and not requiring the use of radioactive isotopes.     

Histopathology and cell culture characteristics of liver cells from grc − and grc + rats given diethylnitrosamine

The histopathological response and cell culture characteristics of liver cells from the R16 (grc ^–) strain of rats, which carries an MHC-linked deletion, were examined one week after a single intraperitoneal injection of 200 mg/ kg body weight diethylnitrosamine (DEN) and were compared with the response of liver cells from wild type (grc+) rats. The DEN exposure induced hydropicl vacuolar changes in the parenchymal cells and a limited proliferation of oval cells in the periportal areas of the livers of both grc+ and grc rats. Primary culture of collagenase-digested livers consisted of parenchymal, bile ductular and oval-related cells as determined by cell-specific immunohistochemistry. Subpassaged cells from grc+ rats exhibited oval cell ultrastructural morphology, inducible histochemical staining for gammaglutamyl transpeptidase (GGT), and DEN-associated onset of anchorage-independent growth. Primary cultures of liver cells from R16 rats consistently failed to form cell strains upon subpassage.Abbreviations DEN diethylnitrosamine - grc growth and reproduction complex - GGT gamma-glutamyl transpeptidase - MHC major histocompatibility complex     

Serum copper,zinc levels,and copper

Serum copper and zinc levels were determined in 20 healthy women and in 100 women with gynecological tumors. Malignant and benign tumor cases were separated according to their postoperative, histopathological examinations. The stages of malignant and benign tumors were also established histologically. Seventy benign and 30 malignant genital tumors (carcinoma of cervix in situ, cervix, ovary endometrium, and vulva) of the patients were differentiated histopathologically. The serum Cu/Zn ratios of patients were increased significantly from the control group (0.32±0.35) to the benign group (1.22±0.63) and from the benign group to the malignant group (2.24±1.03). Nine of 30 malignant cases were determined as false negative (30%) and 15 of 70 benign cases were determined as false positive (14.2%) according to the serum Cu/Zn ratios of patients. Serum copper levels of 30 malignant and 10 benign tumor cases showed linear correlation with serum ceruloplasmin values.     

区别人小细胞肺癌和非小细胞肺癌的LSC系列单克隆抗体(简报)

人小细胞肺癌的发病率虽不及非小细胞肺癌,但其五年生存率却相对低得多,这两种细胞类型不仅在病理类型上表现不同,在许多其它性质上也表现不同。小细胞肺癌是一种异质性疾病,它由几种形态不同的瘤细胞组成,包括非小细胞肺癌在内;它易早期转移,能在肝、脑和骨髓中繁殖,具有L-Dopa胱羧酶,神经原特异的烯醇化酶(enolase),能合成神经递质和多肽激素蛙皮肽等特性。目前已有关于小细胞肺癌的单克隆抗体的报道,我们采用美国N I H建立的小细胞肺癌细胞株SH-77作免疫原,目前尚未见有关抗该细胞株的单克隆抗体的报道,现将我们制备的三个单抗体及其初步特异性鉴定结果简报如下     

钙调素抑制剂——三氟拉嗪对肿瘤细胞增殖和微管组装的影响

本文用钙调素抑制剂——三氟拉嗪处理人胃癌MGC-803细胞,用免疫荧光细胞化学方法,放射免疫法和速流荧光分析等方法研究了钙调素对细胞增殖,环核苷酸代谢及微管组装,有丝分裂等细胞功能的调节作用。实验结果表明,TFP明显地抑制了人胃癌细胞的增殖,这种抑制增殖的作用,具有剂量和时间依赖关系,细胞群体中G_1期细胞增多,S期细胞下降,DNA合成明显地受到抑制。TFP处理的胃癌细胞仅在短时间内(5'-30')cAMP含量升高,cGMP浓度降低,cAMP/ ??cGMP比值比对照组高4.4倍,但此后环核苷酸含量又很快恢复到对照组水平。本实验还观察到TFP处理后的MGC-803细胞胞质铺展,细胞形态的改变与胞质微管的分布有密切联系,实验结果表明TFP加强了人胃癌细胞MTOC对微管的组装能力,使微管分布得到恢复,微管纤维呈放射状延伸到细胞边缘,充满胞浆,使细胞呈现出展平的多边形,趋向于正常上皮细胞形态的变化,本实验结果表明TFP抑制癌细胞增殖及使微管组装加强可能是通过对CaM活性的抑制作用。此结果有助于说明转化细胞内钙调素的变化,可能是与转化细胞增殖失控和胞质微管消退有关。     

Cell surface heparan sulfate proteoglycan and the neoplastic phenotype

Cell surface proteoglycans are strategically positioned to regulate interactions between cells and their surrounding environment. Such interactions play key roles in several biological processes, such as cell recognition, adhesion, migration, and growth. These biological functions are in turn necessary for the maintenance of differentiated phenotype and for normal and neoplastic development. There is ample evidence that a special type of proteoglycan bearing heparan sulfate side chains is localized at the cell surface in a variety of epithelial and mesenchymal cells. This molecule exhibits selective patterns of reactivity with various constituents of the extracellular matrix and plasma membrane, and can act as growth modulator or as a receptor. Certainly, during cell division, membrane constituents undergo profound rearrangement, and proteoglycans may be intimately involved in such processes. The present work will focus on recent advances in our understanding of these complex macromolecules and will attempt to elucidate the biosynthesis, the structural diversity, the modes of cell surface association, and the turnover of heparan sulfate proteoglycans in various cell systems. It will then review the multiple proposed roles of this molecule, with particular emphasis on the binding properties and the interactions with various intracellular and extracellular elements. Finally, it will focus on the alterations associated with the neoplastic phenotype and will discuss the possible consequences that heparan sulfate may have on the growth of normal and transformed cells.     

DNA拓扑异构酶Ⅰ生物学活性的比较研究

测定了3T3细胞、人和大鼠一些组织中DNA拓扑异构酶Ⅰ的活性;估计了核酸内切酶对拓扑酶Ⅰ松弛活性测定的干扰程度;发现增殖组织全细胞抽提液中酶比活高于正常分化组织,而且在异常增殖组织中酶比活的增高更为显著。     

Evaluation of microwave hyperthermia applicators.

Hyperthermia has been used in conjunction with radiation and chemotherapy for cancer treatment. When using electromagnetic heating, applicators are critical components in contact with or in proximity to patients and can be the determining factor for effective and safe treatment. Tissue absorption of electromagnetic energy is determined by many factors. Three cases are shown to illustrate the complexity of microwave heating: 1) The BSD MA-151 applicator has good center heating on a muscle-only phantom as shown in the operation manual. When fat slabs of 0.25, 0.5, 1, and 2 cm thick were added, two hot spots near the periphery of the applicator were evident on all fat surfaces, exposed at 631 MHz. At 915 MHz, the heating was elongated on the surface of the models with 0.25- and 2-cm fat, and two hot spots were observed on the 0.5- and 1-cm fat surfaces. 2) Heating patterns of the Clini-Therm applicators on a muscle-only phantom, as indicated in the operations guide, are elliptical with their major axes perpendicular to the electric field. However, when a bolus is used, the elliptical pattern is parallel to the E field. 3) Heating patterns in cylindrical structures were studied with inhomogeneous models of limbs. Arm and thigh models consisting of fat, bone, and muscle material were heated with Clini-Therm L, M, and MS applicators at 915 MHz. In addition to the geometric effect, the results indicated that placing the applicators with E field parallel to the long axis of cylindrical structures can minimize required power, produce less heating of fats and reduce stray radiation. In conclusion, to apply penetrating microwave or other RF fields for tissue heating, one must simulate the clinical exposure conditions as closely as possible to obtain useful heating patterns.     

Active specific immunotherapy with Newcastle-diseasevirus-modified autologous tumor cells following resection of liver metastases in colorectal cancer

Summary A group of 23 colorectal cancer patients were treated by a new type of active specific immunotherapy (ASI) following complete surgical resection of liver metastases (R0 resection). For ASI treatment we used a vaccine consisting of 1 × 10⁷ autologous, irradiated (200 Gy) metastases-derived tumor cells incubated with 32 hemagglutination units (HU) of Newcastle disease virus (NDV). The adjuvant vaccine therapy was started 2 weeks after surgery and was repeated five times at 14-days intervals followed by one boost 3 months later. The delayed-type hypersensitivity (DTH) skin reactions to the vaccine were measured as well as the DTH reactions to a challenge test of 1 × 10⁷ non-virus-modified autologous tumor cells from liver metastases or 1 × 10⁷ autologous normal liver cells. In addition 32 HU NDV alone and a standard antigen test (Merieux test) were applied pre- and post-vaccination. The vaccination was well tolerated. In 13 of 23 patients an increasing reactivity against the vaccine was observed during the vaccination procedure. Nine patients (40%) experienced an increased DTH reactivity against autologous tumor cells following vaccination, while 17% or fewer showed an increased reactivity to Merieux test antigens, NDV, or normal liver cells. The increased antitumor response was not correlated to responsiveness to NDV alone, autologous liver cells, enzymes and culture medium used for vaccine preparation or standard antigens (Merieux test). After a follow-up of at least 18 months 61% of the vaccinated patients developed tumor recurrence in comparison to 87% of a matched control groups from the same institution that had been only surgically treated. The results of this phase II trial are encouraging and should stimulate further prospective randomized studies.     

Modification of blood group A antigen expression in a pancreatic cancer cell line (PC-1) by inhibitors of N-glycan processing.

Pancreatic adenocarcinomas induced in Syrian hamsters by treatment with N-nitrosobis(2-oxopropyl) amine express blood group A antigen, which is absent in normal pancreatic cells. On membrane glycoproteins purified from tumors, blood group A antigen has been found to be expressed on multiantennary Asn-linked complex glycans. In this study, we investigated the effect of inhibitors of Asn-glycan processing on blood group A antigen bearing glycan structures in a cell line (PC-1) established from a primary induced pancreatic cancer. Expression of blood group A antigen on cells and in membrane preparations was blocked by treatment with 1-deoxymannojirimycin, an inhibitor of mannosidase I, but was retained after treatment with swainsonine, an inhibitor of mannosidase II. However, swainsonine treatment altered the glycan structure associated with blood group A antigen from an endoglycosidase H resistant type to a sensitive type, indicating that the blood group A structure might shift from a complex type to a hybrid type glycan by this treatment. These results demonstrate that Asn-linked glycans carry the major blood group A antigens in PC-1 cells.