U50488 inhibits outwardly rectifying potassium channel in PC12 cells via pertussis toxin-sensitive G-protein

This study was undertaken to determine the effect of U50488, a kappa-opioid receptor agonist, on outwardly rectifying potassium channel (Ik) in undifferentiated PC12 cells. Using whole-cell and on-cell patch-clamp techniques, we found that U50488 decreased Ik amplitude in a time-dependent manner and Ik activation was delayed. Single-channel kinetic analysis provided a two-stage model for us to illuminate the blockage effect induced by U50488. To identify whether U50488 mediates the effect through opioid receptor and G-protein, several specific blockers and activators were used. Not only naloxone but also PTX and GDPbetaS abolished U50488-induced suppression; however, such effect was not observed when cAMP or other adenylyl cyclase activators were used. It is postulated that kappa-opioid receptor and Gi/o protein, but not cAMP, are involved in U50488-induced suppression of Ik.     

百日咳抗体酶联检测试剂盒的研制及其应用

为了研制百日咳抗体酶联检测试剂盒以调查吉林省2~8岁儿童百日咳、白喉及破伤风抗体水平。采用百日咳菌液经硫酸铵盐析、PBS溶液浸提及蔗糖密度梯度离心提取的PT和FHA作为包被抗原,辣根过氧化物酶(HRP)标记羊抗人IgG,作为抗体制备ELISA试剂盒。并经敏感性、特异性、重复性试验考查该试剂盒质量。结果显示,试剂盒敏感性可达0.0036 IU/m l;重复性较好,CV<4%。试剂盒置于37℃3d敏感性无明显变化。吉林省2~8岁儿童白喉、破伤风的抗体水平较高,百日咳的抗体水平相对较低。此方法制备的百日咳抗体酶联检测试剂盒的质量符合要求,可应用于临床检验与流行病学监测。     

Seasonality of Primarily Childhood and Young Adult Infectious Diseases in the United States

The Center for Disease Control (CDC) in the United States collects and maintains records of communicable (so‐called notifiable) infectious diseases that cause significant morbidity and mortality and impact the national economy. This investigation focused on seasonal patterns in the primarily childhood and young adult infectious diseases of meningococcal meningitis, mumps, pertussis, typhoid fever, streptococcal toxic shock syndrome (1990 to 2003 CDC database), and varicella (1993 to 2003 CDC database). Linear regression was performed to ascertain the trend in the incidence of each disease, and multi‐component cosinor analysis was applied to determine and describe periodicities. Significant decreasing trends in incidence were detected in meningococcal meningitis, mumps, typhoid fever, and streptococcal toxic shock syndrome, and increasing trends were found in pertussis and varicella. Significant annual patterns were documented in meningococcal meningitis (January peak), mumps (April peak), pertussis (August peak), varicella (April peak), typhoid fever (August peak), and in the hospital‐acquired streptococcal toxic shock syndrome (February peak). Such seasonal patterns and long‐term trends in infectious diseases are of practical public health significance in indicating which can benefit from timely prevention interventions.     

Insulin stimulates the release of a subset of GPI-anchored proteins in a G-protein independent manner

The glycosyl-phosphatidylinositol anchored protein, membrane dipeptidase (EC 3.4.13.19) is released from the surface of 3T3-L1 adipocytes in response to insulin treatment through the action of a phospholipase C. The present study investigates the role of guanine-nucleotide binding proteins (G-proteins) in this process. Treatment of permeabilized 3T3-L1 adipocytes with GTP gamma S did not cause release of membrane dipeptidase into the medium, while GDP beta S did not inhibit the insulin-stimulated release of membrane dipeptidase. Other activators of G-proteins, including the tetradecapeptide mastoparan, pertussis toxin and AlF, also caused no significant release of membrane dipeptidase from the surface of the 3T3-L1 adipocytes. From these observations it is concluded that G-proteins are not involved in the insulin stimulated release of membrane dipeptidase. Although X-Pro aminopept idase (EC 3.4.11.9) is GPI-anchored in 3T3-L1 adipocytes as shown by digestion with bacterial phosphatidylinositol specific phospholipase C, it was not released upon insulin treatment of the cells, indicating that only a subset of the GPI-anchored proteins are susceptible to insulin-stimulated release.     

平顶山市2004—2011年百日咳流行特征分析

目的了解平顶山市百日咳发病流行规律,为有效防控百日咳提供科学依据。方法采用描述流行病学方法分析,对平顶山市法定传染病报告系统和近年对百日咳疫情监测资料进行分析,对百日咳的时间分布、疫苗效果等进行比较。结果自实施计划免疫以后,百日咳发病率和死亡率大幅度下降,年均发病率由20世纪60～70年代的93.08/10万降至目前的1/10万以下;流行范围逐步缩小,春夏为高发季节,主要集中在5～8月份,病例10岁以下儿童共88例,占病例总数的98.88%,主要集中在7岁以下的为57例,占64.05%。城市高于农村,两者发病率差异有统计学意义。女性多于男性。职业以散居、托幼儿童为主,占发病总人数的57.30%;其次为学生,占41.58%。结论防治百日咳应进一步提高和保持高水平的百白破混合制剂常规免疫接种率,控制局部地区的爆发,并加强监测和传染源的管理。     

中国百日咳疫苗的现状及研发趋势初探

百日咳是许多国家的严重公共卫生问题,目前预防百日咳最有效和最经济的方法是接种百日咳疫苗,百日咳组分疫苗由于其保护效果好、不良反应低、质量稳定可控而成为新一代无细胞百日咳疫苗的首选。本文简要介绍百日咳的病原学、传播途径及流行病学特点,并对中国的百日咳疫苗研发趋势进行初步探讨。     

Production and secretion of pertussis toxin subunits in Bacillus subtilis

Pertussis toxin (PT) is a major component of today's acellular whooping cough vaccines. The use of acellular vaccines is predicted to increase sharply in the near future. There is therefore a need to produce PT in a way that makes its purification as easy as possible. Our approach was to express all five PT subunits individually in Bacillus subtilis. We have used vectors containing the promoter and signal sequences of the alpha-amylase gene of Bacillus amyloliquefaciens followed by an insert encoding the appropriate PT-subunit. All PT-subunits were secreted and found in the culture supernatant. The level of expression varied considerably: S1 and S5 were produced in large quantities whereas much smaller amounts of S2, S3 and S4 were found. The subunits were also present in the membrane fraction of the respective strains.     

μ-Opioid Receptors and Not K-Opioid Receptors Are Coupled to the Adenylate Cyclase in the Cerebellum

The putative regulatory effect of opioids on adenylate cyclase was investigated in two different preparations containing, respectively, two different populations of opioid receptors: the rabbit cerebellum (greater than 75% mu-opioid receptors) and the guinea pig cerebellum (greater than 80% kappa-opioid receptors). In the mu-preparation, but not in the kappa-preparation, opioids inhibited the basal and the forskolin-stimulated adenylate cyclase activity in a dose-dependent manner and stereospecifically. The inhibition was in the 20-30% range, required the presence in the assay medium of Mg2+ and of GTP, but was independent of the presence of Na+. Pharmacological characterization of the inhibitory response in the rabbit cerebellum clearly showed that it was under the control of a mu-opioid binding site, with the effect being elicited by non-selective (etorphine and morphine) and mu-selective (Tyr-D-Ala-Gly-Me-Phe-Gly-ol) agonists, whereas delta- and kappa-selective agonists were almost totally ineffective. ADP ribosylation of inhibitory GTP-binding protein by pertussis toxin failed to block the inhibitory effect of opioids, and data presented suggest that this failure is likely to be the consequence of a limited access of the toxin to its substrate in rabbit cerebellum membranes.     

The inflammatory effects of UDP-glucose in N9 microglia are not mediated by P2Y14 receptor activation

In this study we evaluated the functionality and inflammatory effects of P2Y14 receptors in murine N9 microglia. The selective P2Y14 receptor agonist UDP-glucose (UDPG) derived from microbial sources dose dependently stimulated expression of cyclooxygenase-2 and inducible nitric oxide synthase, and potentiated the effects of bacterial lipopolysaccharide on nitric oxide production. However, another selective P2Y14 receptor agonist, UDP-galactose, did not affect these endpoints either alone or in combination with lipopolysaccharide. Interestingly, synthetic UDPG also had no detectable pro-inflammatory effects, although P2Y14 receptors are both expressed and functional in N9 microglia. While synthetic UDPG decreased levels of phosphorylated cyclic AMP response element binding protein, an effect that was blocked by pertussis toxin, the pro-inflammatory effects of microbial-derived UDPG were insensitive to pertussis toxin. These data suggest that the pro-inflammatory effects of microbial-derived UDPG are independent of P2Y14 receptors and imply that microbial-derived contaminants in the UDPG preparation may be involved in the observed inflammatory effects. An erratum to this article can be found at