Investigation of Role of Nitric Oxide in Protection from Bordetella pertussis Respiratory Challenge

The mechanism whereby whole-cell pertussis vaccines (WCV) confer protection against Bordetella pertussis is still not fully understood. We have previously reported that macrophage activation produced by vaccination with WCV is associated with induction of NO synthesis by macrophages in response to in vitro stimulation with B. pertussis antigens. To determine whether NO production is an effector of protection or simply a marker of activation, the susceptibility of inducible nitric oxide synthase (type II, iNOS) knockout mice to infection with B. pertussis was examined. We showed that iNOS knockout mice were more susceptible to B. pertussis respiratory challenge than wild-type mice. iNOS-deficient mice also developed a less effective protective response than wild-type mice after the same immunization with WCV. This suggests that NO plays an important role in effecting protection against B. pertussis challenge.     

111例双黄连过敏反应文献分析

目的:分析临床应用双黄连引致的过敏反应的规律。方法:对2000年1月-2004年3月中国期刊网中收录的111例双黄连过敏反应在性别、年龄、发生时间、给药途径和临床表现进行分析。结果:双黄连过敏与年龄、性别无关,与给药途径有关,注射剂易发生过敏反应。该药过敏的临床表现有:过敏性休克67例(60%),皮肤黏膜症状61例(55%),其他类型过敏反应14例(13%)31例同时出现多种过敏症状。过敏发生时间最短为开始用药1m in以内,最长为用药后72h,97例(87%)在用药过程中发生过敏症状。结论:双黄连的过敏反应以速发型过敏反应(如过敏性休克)为主,应是临床关注的重点。     

Spermidine synthase of Escherichia coli: localization of the speE gene.

We have obtained Escherichia coli mutants lacking spermidine synthase (putrescine aminopropyltransferase) and have found that the mutated gene (speE) is located immediately upstream from the gene coding for S-adenosylmethionine decarboxylase (speD); these genes are located at 2.7 minutes on the E. coli chromosome. Both genes are present in a 1795-base-pair fragment of E. coli DNA that was cloned into pBR322. Deletion of 105 bases upstream of speE caused a coordinate loss of both activities, indicating that speE and speD constitute a single operon. speE and speD have also been cloned separately in a high-expression vector; strains carrying these plasmids overproduce the respective enzymes.     

Effect of gradual rise in plasma calcium concentration on the impairment of atrioventricular nodal conduction due to verapamil

The possible reversal by calcium of the inhibitory action of verapamil on the atrioventricular (AV) node was investigated in anesthetized, atropinized dogs, with cardiac pacing. The His bundle potentials were recorded by endocavitory electrode and the AV node effective refractory period measured by the extrastimulus method. Calcium infusion was effective against the impairment of AV nodal conduction induced by verapamil, provided it remained moderate: the gradual rise in the plasma calcium concentration counteracted the effects of an infusion of verapamil on conduction time and effective refractory period in the AV node, as long as it did not exceed 5 mmol/L. However, beyond this level, calcium appeared less and less capable of reversing the effects of verapamil. Thus, the protective action of calcium had a bell-shaped dose-response curve, with the optimum at 5 mmol/L. This biphasic influence is consistent with the opposite opinions previously given concerning the antagonism between calcium and calcium blockers, depending on whether hypercalcemia brought into play was mild or major. In any case, the prominent role played by calcium in the slow inward current in the AV node accounts for the antagonism, observed in vivo, between calcium and verapamil. The pacemaker activity of the sinoatrial (SA) node was less influenced by both calcium blocker and calcium.     

Functional recognition of the neuronal tyrosine hydroxylase cAMP regulatory element in different cell types.

Transient transfection of pLB2CAT constructs bearing short synthetic oligonucleotides derived either from the tyrosine hydroxylase (TH) promoter or other sources was used to examine functional cAMP regulatory element (CRE) activity in a variety of cell lines. The region containing only the putative TH CRE was found to be as or more effective in conferring cAMP responsiveness onto pLB2CAT (which employs the TK promoter) than the immediate 272 bp region of the TH promoter. Increases in CAT activity of 10- to 20-fold were observed in JEG-3 cells with a single insert of the TH CRE region (-31 to -54) in pLB2CAT, and the presence of a second insert generated only a modest further increase. This construct also responded to cAMP in 4 other cell lines tested but the degree of increase was less dramatic. Inserts containing the consensus 8 bp CRE motif embedded in other natural or artificial contexts served generally as weak functional CREs in all cell lines tested. In vitro analysis revealed that a specific protein-DNA complex apparently containing a single protein with a MW of 45-50 kDa was formed equally well with JEG-3 cell nuclear extract and CRE-bearing-TH and other fragments which produced dramatically different cAMP effects in vivo. These results suggest specificity in the effects of cAMP on different CREs which are dictated by contextual differences.     

二乙基亚硝胺诱导大鼠肝细胞癌过程中明胶酶的动态变化

目的：观察肝癌形成过程中明胶酶表达（蛋白及mRNA水平）的动态变化，探索明胶酶在肝癌生长浸润中的作用。方法：应用免疫组化法、明胶酶谱法和RT-PCR法对大鼠肝癌形成过程中各期明胶酶表达情况进行观察，并与正常对照组进行比较分析。结果：正常肝组织及肝硬化组织内明胶酶有表达，肝癌形成后主要在癌细胞内表达；明胶酶原在正常肝组织中有低表达，诱癌过程中呈持续增高趋势；明胶酶-2mRNA与其酶蛋白表达趋向一致。结论：在肝癌形成过程中明胶酶转录和翻译水平均呈持续增高趋势。     

捻转补泻手法针刺合谷穴对其局部皮肤温度的影响

目的阐明针刺捻转补法与泻法的操作是否存在效应上的差异,并探讨其效应差异是否为补泻效应的差异.方法应用红外线热像技术,采用不同的捻转补泻手法针刺健康人合谷穴后,观察其在即刻,10、20、30min,对局部皮肤温度的影响.结果不同捻转补法与泻法的操作存在着不同程度的效应差异,其中补法可以使皮温升高,泻法可以使皮温降低,以石氏捻转补泻针法较为明显.结论1)补泻手法,补法和泻法的操作可产生不同的效应.2)不同捻转补泻手法对皮肤温度产生的升降效应为补泻效应,其中以石氏捻转补泻手法最为明显.3)证明补泻手法实施的必要性.     

Calcium-activated proteolysis of intracellular domains in the cell adhesion molecules NCAM and N-cadherin.

One of the consequences of increased intracellular calcium in response to a variety of physiological stimuli is the calcium activation of cytosolic proteases. Unlike lysosomal proteases with broad specificity, these calcium-activated neutral proteases show limited proteolysis of a restricted set of substrate proteins suggesting they may play a regulatory role in cellular physiology. In this study we show that the neural cell adhesion molecules NCAM-180 and N-cadherin are substrates for such endogenous calcium-activated neutral proteases. In contrast, a third neural cell adhesion molecule G4/L1 was not susceptible to calcium-activated proteolysis. The threshold for activation of NCAM and N-cadherin proteolysis is in the micromolar range of calcium suggesting that NCAM and N-cadherin are substrates for a mu-type calpain (calpain I). The site recognized by this protease is within intracellular domains of NCAM-180 and N-cadherin which are important for their interaction with cytoskeletal components. These results suggest that calcium-activated proteolysis at these sites in vivo could disrupt the linkage between extracellular ligand binding to these adhesion molecules and the normal intracellular effectors of such extracellular binding events.