A sensitive sandwich ELISA for measuring thrombopoietin in human serum: serum thrombopoietin levels in healthy volunteers and in patients with haemopoietic disorders

A sensitive sandwich enzyme-linked immunosorbent assay (ELISA) has been established to estimate serum thrombopoietin (TPO) concentrations in healthy volunteers and patients with haemopoietic disorders. The ELISA uses a mouse monoclonal antibody (Ab) as the capture Ab and a biotinylated rabbit polyclonal Ab as the detector. The ELISA was reproducible, highly sensitive and specific for human TPO. The coefficients of intra- and inter-assay variation were from 3.0% to 4.9% and from 5.9% to 6.1%, respectively. The quantitative limit of the ELISA was 0.09 fmol/ml in serum. The quantitative limit was lower than the normal level. The dose–response curves of serum samples from healthy volunteers and patients with haemopoietic disorders were parallel to the standard curves. The ELISA did not cross-react with a variety of blood components and cytokines to produce false-positive results.
The serum TPO concentrations from 29 normal males and 21 females were 0.79 ± 0.35 and 0.70 ± 0.26 fmol/ml, respectively. Serum TPO levels in patients with aplastic anaemia (AA), acute lymphocytic leukaemia (ALL) and essential thrombocythaemia (ET) were measured using the ELISA. The serum TPO levels in the patients with ET ( n  = 6, 2.80 ± 1.55 fmol/ml) were higher than the normal level. The patients with AA ( n  = 7, 18.53 ± 12.37 fmol/ml) and ALL ( n  = 5, 10.36 ± 5.57 fmol/ml) had significantly higher serum TPO levels than normal individuals. These results indicate that the ELISA specific to TPO should prove useful in measuring the TPO concentration in serum samples.     

甲磺酸伊马替尼抑制黑素细胞游走的实验研究

目的探讨甲磺酸伊马替尼对人黑素细胞(NHEM)游走的抑制作用。方法培养NHEM,用四甲基偶氮唑蓝(MTT)还原法测定甲磺酸伊马替尼对NHEM生长和分化的影响;使用侵袭小室测定甲磺酸伊马替尼抑制NHEM的游走情况;用蛋白印记法测定甲磺酸伊马替尼抑制c-kit磷酸化情况。结果甲磺酸伊马替尼0.1μmol/L,1.0μmol/L,5.0μmol/L能够抑制由SCF50ng/mL引起的NHEM游走,并能抑制c-kit的磷酸化。结论甲磺酸伊马替尼抑制SCF引起的NHEM游走及抑制c-kit的磷酸化,为其用于防治色素痣治疗后的复发奠定了理论基础。     

International cooperation with Islamabad Children's Hospital,Pakistan: The JICA project

The experience gained during 7 years of cooperation between the Japan International Cooperation Agency (JICA) and the Islamabad Children's Hospital (JICA-ICH project, July 1986-June 1993) is described. Islamabad Children's Hospital achieved the goals of the project and became a centre for excellence in health care, education and research for children, fulfilling the objectives of the project. This achievement was evaluated as one of the most successful projects in medical cooperation ever performed by JICA by a third party evaluation team. The problems arising and the lessons experienced through the process are discussed. The importance of the role which should be undertaken by pediatricians in international cooperation with developing countries is emphasized.     

Three Cases of Esophageal Granular Cell Tumor

Abstract: Although it has long been thought that granular cell tumor (GCT) is relatively uncommon in the esophagus, in recent years, reports of this disease have increased due to advances in endoscopic examination and endoscopic therapy. The authors recently experienced three cases of esophageal GCT, all of whom underwent endoscopic polypectomy. Endoscopic findings were consistent with Yamada's type I or II, the surface of the lesions being smooth and the color white or whitish-yellow. These three cases were treated by endoscopic polypectomy. In case 1, the resection was made possible by raising the tumor with forceps under a 2-channel-scope. In case 2, the tumor was resectable following submucosal injection of physiological saline. In case 3, the tumor was resected via strangulation with a snare. The lesions described herein were diagnosed as benign and completely resected by polypectomy, though some showed differences in nuclear size or dyskaryosis. As numerous points remain to be clarified regarding the clinical characteristics of this tumor, and some tumors have been diagnosed as malignant despite being small, it appears that endoscopic polypectomy should be performed for the purpose of diagnosis as well as complete resection.     

Differential expression of phosphorylated extracellular signal-regulated kinase 1/2, phosphorylated p38 mitogen-activated protein kinase and nuclear factor-κB p105/p50 in chronic inflammatory skin diseases

The keratinocytes actively participate in the cutaneous immune responses. Dysregulation and abnormal expression of inflammatory mediators or their receptors in keratinocytes are relevant to the pathogenesis of chronic inflammatory skin diseases. The mechanism of long-lasting inflammatory processes is related with the activation of nuclear factor (NF)-κB and mitogen-activated protein kinase (MAPK), which play a crucial role in the immune responses. There are potential interaction points between these two pathways. The aim of this study is to investigate the differences in expression levels and distributions of phosphorylated extracellular signal-regulated kinase (ERK)1/2, phosphorylated p38 MAPK and NF-κB p105/p50 in chronic inflammatory skin diseases. An immunohistochemical staining technique was employed to measure the expression of these molecules in 25 cases of lichen planus, 22 cases of psoriasis, 26 cases of chronic eczema, seven cases of prurigo and seven cases of normal skin. We observed that the expression of phosphorylated ERK1/2, phosphorylated p38 MAPK and NF-κB p105/p50 was significantly more augmented in the lesional epidermis of all the inflammatory skin diseases than those in normal skin ( P  < 0.05), and the number of positive keratinocytes was significantly more in lichen planus than that in other inflammatory diseases ( P  < 0.001). Moreover, the positive keratinocytes of these three molecules were more widely distributed in the entire layer of the epidermis in lichen planus than those in other diseases. We concluded that ERK1/2, p38 MAPK and NF-κB p105/p50 might play important roles in the pathophysiology of chronic inflammatory skin diseases.     

Gastric Cancer Developing in a Patient with Chronic Lymphocytic Leukemia with Massive Gastrointestinal Involvement

Massive infiltration of neoplastic lymphocytes caused a variety of gross gastrointestinal manifestations in a 71-year-old patient with chronic lymphocytic leukemia (CLL). These included plaque-like elevations in the esophagus, convoluted gyrus-like folds in the stomach, and many polypoid lesions in the stomach and duodenum. No gross lesions were noted in the large intestine. Four years later, systemic chemotherapy reduced the gastrointestinal involvement, but a papillary elevation grew at the gastric angle, which was shown to be an early cancer on histological examination. Gastrointestinal involvement in CLL and second cancers associated with CLL are reviewed, with special reference to the possible relationship between CLL and gastric cancer.     

Evaluation of Esophago-gastric Varices with Endoscopic Ultrasonography

Esophago-gastric varices of 22 patients were studied using a newly developed method of endoscopic ultrasonography (EUS). During the observation, the esophagus was filled with de-aerated water, and reflux of the water was prevented using a balloon placed 7 cm proximal to the tip of the endoscope. Thirteen of 22 patients received endoscopic sclerotherapy (EIS) for their esophago-gastric varices, and sequential changes of the varices were observed mith EUS. The EUS method demonstrated esophago-gastric varices as singular, or a bundle of low-echoic luminal structures, and extramural collateral vessels were also observed. Therefore, it was possible to evaluate the residual blood flow after EIS by the EUS findings. Sequential changes of the treated varices were observed as follows: (1) formation of a thrombus in the varices, (2) thickening of variceal wall, and (3) disappearance of luminal structures. Small luminal structures in the extramural space of the lower esophagus, dilated paraesophageal veins, were detected in 6 of 13 patients with EIS. The recurrence rate was smaller and the remission period was longer in these 6 patients than those in patients without these luminal structures. Our new EUS method has been shown to be useful not only for the morphological evaluation of varices, but also for the assessment of the effect of EIS.     

p38 MAPK/NF-κB/cyclin D1信号传导通路在乳房外Paget's病中的意义

目的探讨磷酸化p38细胞丝裂原活化蛋白激酶α(phospho-p38 mitogen-activated protein kinase,p-p38 MAPKα)、磷酸化核因子-κB(phosphor-nuclear factor kappaB,p-NF-κB)及细胞周期蛋白D1(cyclin D1)在乳房外Paget’s病(extramammary Paget’s disease,EMPD)表达的意义及评价它们之间的相关性。方法采用免疫组化ABC法检测了p-p38MAPK,p-NF-κB及Cyclin D1蛋白在来源于30例患者的35张EMPD石蜡包埋切片中的表达(5张来源于有转移的EMPD淋巴结标本)。结果①在35张标本中EMPD石蜡包埋切片中有30例p-p38 MAPK表达阳性,28例p-NF-κB表达阳性,27例cyclin D1表达阳性,并且在其中5张来源于有转移的EMPD淋巴结标本中,5例均p-p38 MAPK和p-NF-κB表达阳性,4例cyclin D1表达阳性;②在EMPD中,p-p38 MAPK,p-NF-κB和cyclin D1阳性表达两两间均有明显的相关性。结论p-p38 MAPK,p-NF-κB和cyclin D1在EMPD中有过度表达,p38 MAPK/NF-κB/cyclin D1信号传导通路可能在EMPD肿瘤形成机制中起重要作用。