人T淋巴母细胞（Jurkat）移动性的研究

利用人Ｔ淋巴母细胞（Ｊｕｒｋａｔ）细胞系作为研究对象。企图探探索细胞外基质成份及肌醇磷脂细胞信息传递系统与细胞移动的关系。研究结果表明：当细胞被伏波醇酯（ＰＭＡ）处理过后，其粘连性与移动件对纤维连结蛋白（ＦＮ）底物的反应在已是高水平的基础上有更进一步提高的作用。与ＦＮ刊相反的是，未经ＰＭＡ处理处的Ｊｕｒｋａｔ细胞对层粘连蛋白（Ｌｍ）底物却无明显的粘连性及移动性增加的反应，尽管在ＰＭＡ处理过４小时内亦无任何的增强。但是，当ＰＭＡ作用于细胞２４一４８小时后则明显池增加其Ｌｍ底物的粘连性及移动性反应，表明了其明显的协同作用。分别用抗ＦＮ、抗Ｌｍ及蛋白激酶Ｃ抑制剂Ｓｔａｕｒｏｓｐｒｉｎｅ（ＳＰ）时均起到特异性的抑制作用。可见，协同作用的产生是通过蛋白激酶Ｃ激活后的－段时间内，使细胞表面Ｌｍ受体密度增加而对底物Ｌｍ反应增强所致。     

膳食纤维与不同蛋白质联用对大鼠胆固醇代谢的影响

观察４种膳食纤维在两种蛋白来源下，对高脂血症大鼠生长、血脂、肝胆固醇、粪类固醇及胆汁成分的影响。结果表明：①四种膳食纤维均可降低大鼠血清ＴＣ水平，尤以豆胶效果显著。②豆胶显著升高ＨＤＬ－Ｃ／ＬＤＬ－Ｃ。膳食蛋白为大豆蛋白时，豆胶、沙棘皮可显著升高血清ＨＤＬ－Ｃ、ＨＤＬ－Ｃ／ＴＣ。③豆胶显著减少肝胆固醇累积，两种沙棘皮与大豆蛋白联用降脂强于与酪蛋白共用。④蛋白来源显著影响粪类固醇的排出，纤维类型可影响中性固醇排出并与蛋白质有交互作用。提示：蛋白质与膳食纤维之间的交互作用是探讨营养素与脂质代谢不可忽视的因素。     

Fibrinogen promotes monocyte adhesion via a protein kinase C dependent mechanism

The accumulation of blood monocytes at sites of predilection of the vessel wall is an early cellular event of atherogenesis. Proteins of the vessel wall may facilitate monocyte adhesion and thus promote their recruitment. It has been shown that the relative content of extracellular fibrinogen increases during lesion development, and this study investigated the contribution of immobilized fibrinogen to monocyte adhesion and the underlying mechanism. Freshly isolated human blood monocytes were cultivated in serum-free RPMI 1640 in tissue culture wells precoated with albumin, fibrinogen, or fibrin. After 16 h the plates were washed and adherent cells enumcrated. Immobilized fibrinogen enhanced monocyte adhesion more than 1.9-fold compared to immobilized albumin or fibrin (P<0.05). Concomitant addition of the protein kinase C (PKC) inhibitors staurosporine or H7 suppressed monocyte adherence to immobilized fibrinogen but exerted no significant effect upon adhesion to any other surface tested. Stimulation of monocytes using phorbol myristate acetate resulted in increased binding of monocytes on fibrinogen but not on bovine serum albumin. When PKC activity was reduced through prolonged incubation with PMA for 16h, a significant reduction of monocyte adhesion on fibrinogen was observed. Peptides containing RGD sequences, which have been demonstrated to be ligands for certain integrins, did not inhibit monocyte adhesion. The data suggest that fibrinogen promotes monocyte adhesion in vitro by a PKC-dependent mechanism. PKC appears to be important not only for the initial cell adhesion but also for sustained binding of monocytes to fibrinogen.Abbreviations BSA Bovine serum albumin - ECM Extracellular matrix - PKC Protein kinase C - PMA Phorbol myristate acetate     

环磷酰胺冲击与小剂量持续应用对大鼠性激素影响的比较研究

Wistar大鼠随机分3组，每天组予环磷酰胺组（CP）8mg／kg·d×4W，ip；冲击组予CP56mg／kg·d×4W，ip；对照组予生理盐水1ml／d×4W，ip。结果：①睾丸生精细胞损伤：每日组重于冲击组；②血睾酮（T）含量每日组显著低于冲击组（P＜0．05）；冲击组与对照组无显著差异（P＞0．05）。③对睾丸间质细胞的影响：冲击组轻于每日组；④血FSH、LH含量与垂体重量：每日组与冲击组皆正常；⑤血BUN、Cr水平：3组皆正常．说明CP冲击疗法对睾丸间质细胞损伤较轻；CP除直接损伤睾丸组织外，可能还对下丘脑—垂体—睾丸轴有抑制作用．     

Stimulation of spontaneous transmitter release at the frog neuromuscular junction by 12-O-tetradecanoylphorbol-13-acetate occurs in the absence of extracellular Ca2+ and is enhanced by depolarization

Summary The effect of the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) on the release of transmitter at the frog neuromuscular junction has been investigated electrophysiologically. TPA (100 nmol/l) caused a gradual rise in miniature end-plate potential (MEPP) frequency. After 20–30 min MEPP frequency had risen by approximately 40%. This action of the drug was not inhibited by bathing preparations in either Ca²⁺-free medium (0 Ca²⁺-1 mmol/l EGTA) or high Mg²⁺ medium, or by pretreatment with verapamil (5 mol/l). The inactive TPA analogue 4--TPA had no effect on release rate. There was no indication of any positive correlation between resting MEPP frequency and the size of the subsequent response to TPA treatment. Any synergism between [Ca²⁺]_i and TPA treatment is therefore likely to occur at a site other than that which determines spontaneous release rate.The stimulatory effect of TPA was enhanced 2-fold by carrying out the experiments in a partially depolarising saline (10 mmol/l K⁺). When TPA was applied to preparations bathed in Ca²⁺-free depolarising saline, the response to the drug was still significantly greater than that in non-depolarised preparations. It is concluded that responsiveness to TPA is enhanced by depolarisation, but that little, if any, of this enhancement can be attributed to the consequent influx of Ca²⁺.Send offprint requests to S. J. Publicover at the above addressPEL was in receipt of an S.E.R.C. postgraduate awardZYS was in receipt of financial support from Umm Al Qura University, Saudi Arabia     

Phorbol ester does not mimic,but antagonizes the alpha-adrenoceptor-mediated positive inotropic effect in the rabbit papillary muscle

Summary The phorbol ester 12-O-tetradecanoyl phorbol-13-acetate (TPA) was used to examine the hypothesis that phosphoinositide turnover is involved in the regulation of myocardial contractility mediated by stimulation of alpha-adrenoceptors in the mammalian cardiac muscle. Exposure of the isolated rabbit papillary muscle electrically driven at a rate of 1 Hz at a temperature of 37°C to TPA in concentrations of 10–1000 nmol/l for 30 min did not affect the basal force of contraction. The concentration-response curve for the positive inotropic effect of (–)-phenylephrine mediated by stimulation of alpha-adrenoceptors in the presence of (±)-bupranolol (100 nmol/1) was shifted to the right and downward by TPA in concentrations of 30–1000 nmol/l, while the effect of (–)-phenylephrine mediated by stimulation of beta-adrenoceptors in the presence of prazosin (100 nmol/l) was not decreased, but slightly enhanced by exposure of the muscle to relatively low concentrations of TPA (10–100 nmol/l). Incubation of the membrane fraction isolated from the rabbit ventricular muscle with TPA in vitro under the same condition as employed in the physiological experiments decreased the specific binding of [³H]prazosin but not that of [³H]CGP-12177, while the non-tumor promoting phorbol ester, PDD, was ineffective. These results indicate that activation of protein kinase C by TPA does not mimic the positive inotropic effect of catecholamines mediated by activation of myocardial alpha-adrenoceptors. on the other hand, the specific interaction of alpha-adrenoceptor-mediated processes with TPA in the rabbit papillary muscle is in line with the view that the facilitation of phosphoinositide turnover and subsequent activation of protein kinase C may play a certain role in the coupling of alpha-adrenoceptor occupation by agonists to the process leading to the positive inotropic action. Send offprint requests to M. Endoh     

蛋白质中两种Strand-Loop-Strand模体的判断

目的:蛋白质的结构决定其功能,为了解蛋白质功能,需要了解蛋白质结构。随着基因测序的发展,大量蛋白质一级结构被测出,但直接从蛋白质的序列出发来预测高级结构,仍很困难,尤其是三级结构的预测。然而由超二级结构获得的结构信息可用于三级结构的预测,对蛋白质三维结构及功能预测有重大意义。方法:由一级结构出发,考虑氨基酸序列信息以及氨基酸的亲疏水性,利用Fisher判别法区分两种Strand-Loop-Strand超二级结构模体。结果:采用7交叉检验,Loop长为2～8时,最后平均结果为Q=71.8%,Q_L=68.2%,Q_H=73.4%,MCC=0.39。如Loop长度相差不大时,预测结果更好,如选取Loop长为2、3、4的序列,7交叉检验结果为Q=75.2%,Q_L=69.4%,Q_H=77.7%,MCC=0.45。结论:以氨基酸信息为特征指标,利用Fisher判别法能较好地区分两种Strand-Loop-Strand超二级结构模体。     

上调Mg2+/Mn2+依赖性蛋白磷酸酶1F表达对鼻咽癌HONE-1细胞增殖和迁移的影响

目的 探讨上调Mg²⁺/Mn²⁺依赖性蛋白磷酸酶1F（PPM1F）对鼻咽癌HONE-1细胞增殖、迁移的影响,并阐明其作用机制。 方法 鼻咽癌HONE-1细胞分为pcDNA3.1组（转染pcDNA3.1质粒）和pcDNA3.1-Flag-PPM1F组（转染pcDNA3.1-Flag-PPM1F质粒）。实时荧光定量PCR（RT-qPCR）法和Western blotting法检测2组细胞中PPM1F和E-钙黏蛋白（E-cadherin）mRNA及蛋白表达水平,CCK-8法和克隆形成实验检测2组细胞增殖活性和克隆形成率,细胞划痕实验检测2组细胞划痕愈合率,Transwell实验检测2组细胞中迁移细胞数。 结果 与pcDNA3.1组比较,pcDNA3.1-Flag-PPM1F 组细胞中PPM1F mRNA表达水平明显升高（P<0.01）,且PPM1F蛋白表达量明显增加,细胞中E-cadherin mRNA和蛋白表达水平明显升高（P<0.01）,细胞增殖活性、克隆形成率和划痕愈合率明显降低（P<0.05或P<0.01）,迁移细胞数明显减少（P<0.05）。 结论 上调PPM1F表达能够抑制鼻咽癌HONE-1 细胞增殖和迁移,其机制可能与细胞间的黏附作用有关。     

Diffuse type of senile plaques in the cerebellum of Alzheimer-type dementia demonstrated byβ protein immunostain

Summary We studied senile plaques (SP) in the cerebella of six autopsied subjects with Alzheimer-type dementia (ATD) and ten non-ATD autopsied subjects between the ages of 78 and 90. Neither SP nor amyloid angiopathy (AA) was observed in any of the non-ATD subjects. In the four of the six ATD subjects, diffuse plaques in the molecular layer were seen as ill-defined areas of fine fibrillar materials by protein immunostaining with formic acid pretreatment, the modified Bielschowsky stain, and periodic acid-methenamine silver (PAM) stain. The plaques were not visible with Bodian, Congo red, or periodic acid-Schiff stains. Compact plaques in the Purkinje cell or in the granular cell layers were found in three of the six subjects. Their amyloid core was often surrounded by areolar amyloid deposits. AA was observed in three of the six subjects. The argyrophilia of the diffuse and compact plaques, demonstrated by the modified Bielschowsky and PAM stains, became undetectable when the sections were first treated with formic acid. Such treatment made the plaques immunoreactive with protein antiserum. The findings suggested that cerebellar diffuse plaques and compact plaques consist mainly of an amyloid component, and are characteristic of ATD.     

Connective tissue metabolism in children with chronic renal failure

To assess the characteristics of connective tissue metabolism in chronic renal failure (CRF), urinary excretion of glycosaminoglycan (GAG) fractions and hydroxyproline (HYP) was determined in ten patients with CRF and in ten age-matched healthy children. CRF was found to be associated with elevated free HYP (19.9±6.1 vs 9.8±3.6 mol/day,P<0.05) and depressed peptide HYP excretion (33.1±13.5 vs 225.2±17.7 mol/day,P<0.01), a low rate of total GAG excretion (7.0±2.4 vs 16.1±1.9 mol uronic acid/day,P<0.05) with low chondroitin 4 — sulphate + chondroitin 6 — sulphate (Ch-Ss) (14.0±9.9 vs 65.0±22.1%) and a high proportion of non-sulphated or under-sulphated fractions, i.e. hyaluronic acid + chondroitin + heparan sulphate (HA+Ch+HS) (75.3±11.4 vs 31.5±5.7%). Urinary 3-methyl-histidine (3-met-HIS) excretion and plasma essential free amino acids did not differ in the two groups. In response to haemodialysis no consistent change occurred in urinary excretion of 3-met-HIS, peptide-bound HYP, total GAG or percentage distribution of individual GAG fractions. After haemodialysis all plasma amino acids decreased significantly, and there was a significant increase in urinary excretion of free HYP (P<0.05). We conclude that the alterations in urinary excretion of total and individual GAGs observed in CRF may reflect disturbed connective tissue metabolism which does not appear to be accounted for by protein malnutrition or enhanced protein breakdown and remains uninfluenced by haemodialysis therapy.