Protein phosphorylation in neutrophils from patients with p67-phox- deficient chronic granulomatous disease

Neutrophils are known to contain a major 67-kD protein that undergoes enhanced phosphorylation and translocation to the membrane during cell stimulation. Recent studies have assumed that this 67-kD phosphoprotein is the 67-kD subunit of the phagocyte oxidase (p67-phox). We compare here the protein phosphorylation patterns in lysates of normal neutrophils and neutrophils from patients with chronic granulomatous disease (CGD) that are completely deficient in p67-phox. The phosphoproteins were labeled by incubation of the cells with radioactive inorganic phosphate (32Pi) or by the addition of [gamma- 32P]ATP to electropermeabilized neutrophils. With either method, stimulation of the normal or CGD cells always resulted in an enhanced incorporation of 32p into two proteins in the 67-kD area. The extent of phosphorylation of these two proteins was very similar in the normal and CGD cells when permeabilized neutrophils loaded with [gamma - 32P]ATP were compared. Moreover, no overall differences in the protein phosphorylation patterns were observed between the normal and CGD cells. Our data indicate that the major 67-kD phosphoproteins observed in stimulated neutrophils are clearly different from p67-phox.     

Influence of the calcium ionophores A23187 and X537A on calcitonin secretion from the isolated perfused porcine thyroid

The present study examined the influence of the calcium ionophores A23187 and X537A on the calcitonin (CT) secretory process. The isolated perfused porcine thyroid was used to evaluate ionophore effects on CT secretion and thyroid slices were used to measure 45Ca uptake. Both A23187 and X537A enhanced the rate of CT release from the perfused thyroid. A23187 at a concentration of 19 microM (10 micrograms/ml) produced a maximal increase in CT secretion of 325% above control levels. X537A at a concentration of 16 microM (10 micrograms/ml) produced a peak rise in CT release of approximately 2000% over control levels. The CT secretory response to A23187 was found to be completely calcium dependent; however, the secretory response to X537A was partially, but not completely dependent upon the presence of perfusate calcium. The results demonstrate that these calcium ionophores are very potent CT secretagogues which vary considerably in their calcium dependency.     

Intrauterine growth retardation: evaluation by magnetic resonance. Work in progress

Eleven high risk fetuses between 32 and 37 menstrual weeks gestational age were examined by magnetic resonance (MR) imaging. Serial obstetrical sonograms, birth weights, and serial postnatal examinations were obtained in all subjects. Sagittal MR spin echo images obtained using TR = 0.5 sec and TE = 28 msec were useful for assessing subcutaneous fat. Prospective estimates of fetal fat stores correlated with neonatal outcome better than sonographic measurements of fetal growth parameters or actual birth weight. MR appears to be a safe and useful technique that offers information complementary to obstetrical sonography when IUGR is suspected.     

CT模拟结肠镜在结肠病灶诊断中的应用

结肠癌是常见的消化道肿瘤,上海等地区的发病率有明显增高的趋势。在发达国家,结肠癌占肿瘤死亡率第二位,且与大肠腺瘤关系密切。如能早期发现有恶变先兆的息肉并切除之,可以预防结肠癌发生。目前,对结肠癌发病高危人群并未作大规模普查,部分由于患者缺乏早期主诉,或是由于现有普查手段不够有效。大便隐血试验只能发现３０％～４０％结肠肿瘤,乙状结肠镜不能进入近端结肠,并有１０％～１５％乙状结肠肿瘤漏诊。钡剂灌肠和结肠镜能检查全结肠,但有１０％～１５％患者行结肠镜检查失败,并且结肠镜可能使１０％～２０％病灶遗漏,尚…     

Keratinocyte growth factor-induced motility of breast cancer cells

Endogenous growth factors and cytokines are known to have a major influence on the progression, motility and invasiveness of tumor cells. We have reported previously that conditioned media from mouse fibroblasts increases the motility of breast cancer cells. Further, we determined that keratinocyte growth factor (KGF) was an active factor from mouse fibroblasts responsible for most of the motility response in breast cancer cells. The present study examined the effect of human KGF on the motility of estrogen receptor (ER)-positive and ER-negative human breast cancer cell lines in culture using time-lapse videomicroscopy to quantify cell motility. In the present study we observed that recombinant human KGF enhanced several parameters of cellular motility in ER-positive cells but not in ER-negative cell lines. Further, we observed that the level of KGF receptor (KGFR) expression in ER-positive cells was much greater than in the ER-negative cell lines. The motility response to KGF was found to be both dose-and time-dependent. Of the three ER-positive breast cancer cell lines tested, MCF-7 cells were the most responsive to KGF stimulation. Finally, MCF-7 cells grown in estrogen-depleted media did not respond to KGF. These results suggest that KGF from stromal tissue surrounding a primary tumor mass can enhance tumor cell motility and may be an early signal in the progression of breast cancer cells to a more motile and metastatic phenotype. Thus, KGF, KGFR and/or the KGF signaling pathway may be important therapeutic targets for the treatment or prevention of breast cancer metastasis. This revised version was published online in July 2006 with corrections to the Cover Date.     

中药癌痛克对人肝癌细胞HepG2增殖、凋亡及Rb基因表达的影响

目的:通过观察不同浓度癌痛克对肝癌细胞株HepG2增殖及凋亡的作用,以及在相应状态下细胞内Rb基因表达量的改变,探讨中药癌痛克抗肝癌的可能作用机制。方法:实验于2005-09/2006-03在广州医学院中心实验室完成。取对数生长期的HepG2细胞,使细胞静止于G0/G1期。随机分为癌痛克2,10,50mg/L组和对照组,癌痛克2,10,50mg/L组分别加入对应浓度癌痛克(购自河南省肿瘤研究所,由金蝎、土元、九香虫、大黄、人参、灵芝、黄芪等纯中药组成的粉状制剂,功能:消癌肿、消癌痛、消积水、升白排毒),对照组不加药物,每组设4个复孔。MTT法检测各组细胞24,48,72h增殖率,细胞增殖率=[(A570癌痛克组-A570对照组)/A570对照组]×100%;以流式细胞术检测各组细胞24,48,72h凋亡率;RT-PCR方法检测各组细胞24,48,72h细胞内Rb基因mRNA表达量。结果:①2～50mg/L癌痛克具有明显的增殖抑制作用,癌痛克2,10,50mg/L组在24,48,72h与对照组比较,差异有显著性意义(P<0.05);3组各时点两两之间比较差异有显著性意义(P<0.05);同组各时点之间比较差异有显著性意义(P<0.05)。②2～50mg/L癌痛克组可诱导HepG2细胞凋亡,相同作用时间癌痛克2,10,50mg/L组与对照组比较差异有显著性意义(P<0.001);同一时点各组比较差异有显著性意义(P<0.05);同组不同作用时间点比较差异有显著性意义(P<0.001)。③2～50mg/L癌痛克可上调Rb基因的表达,各浓度组在各时点与对照组比较,差异均有显著性意义(P<0.05);24,48,72h癌痛克2,10,50mg/L组之间比较差异无显著性意义(P>0.05);3组各时点比较差异有显著性意义(P<0.05)。结论:中药癌痛克可通过抑制HepG2细胞增殖或诱导其凋亡,而发挥抗肝癌作用;且其诱导细胞凋亡的机制之一可能为通过增加Rb基因的表达而实现。