真核表达人呼吸道合胞病毒F蛋白的纯化方法

目的 真核表达人呼吸道合胞病毒(human respiratory syncytial virus,SV)融合蛋白(fusion protein,),并完成蛋白纯化及纯度测定.方法 根据编码F蛋白的基因序列设计引物,CR方法扩增出3'端带His标签的F基因序列,克隆入pGEM-T-easy载体,经核酸序列分析后,进一步克隆到pcDNA3.1( )真核表达载体,限制性内切酶鉴定,用脂质体Lipofectamine2000转染COS-7细胞,2 h后再用Westem blot检测目的蛋白的表达.Ni柱亲和层析纯化COS-7细胞表达的F蛋白,高效毛细管电泳分析纯化后蛋白纯度.结果 核酸序列分析证实获得带His标签的RSV F基因序列,没有发生无义突变.转染COS-7细胞后,利用Western blot方法检测到F蛋白的特异性条带,纯度达99%以上.结论 初步建立了真核表达RSV F蛋白的纯化方法,为进一步优化RSV F蛋白制备条件及单克隆抗体及诊断试剂等研究奠定了基础.     

Modulation of agonist binding by guanine nucleotides in CHO cells expressing muscarinic m1 – m5 receptors

In membranes prepared from CHO-m2 cells, inhibition of [³H]-N-methylscopolamine ([³H]NMS) binding by several muscarinic agonists resulted in competition curves with Hill slopes significantly different from unity. Addition of 5-guanylylimidodiphosphate (Gpp(NH)p) led to an increase in the IC₅₀ value of the agonists with significant steepening of the inhibition curves. The shift in potency induced by Gpp(NH)p differed among the agonists with a rank order of oxotremorine-M = carbachol > oxotremorine > McN-A-343 = pilocarpine. In CHO-m4 membranes, Gpp(NH)p was less efficacious than in CHO-m2 membranes whereas no effect of the guanine nucleotide was found in membranes prepared from CHO-m1, -m3, and-m5 cells. No major differences in the effect of Gpp(NH)p among agonists were found in CHO-m4 cells. Atropine binding was not affected by the guanine nucleotide. Together, these results indicate that coupling of G-proteins to muscarinic receptors linked to inhibition of cyclic adenosine monophosphate (cAMP) (m2 and m4) but not of those linked to phosphoinositol turnover (m1, m3 and m5) can be perturbed by Gpp(NH)p. The differential effects observed with Gpp(NH)p between agonist binding to m2 and m4 receptors appear to be receptor-specific and may reflect differences in the G proteins activated by these receptors in CHO cells.     

Safety of tiagabine: summary of 53 trials

We reviewed the clinical safety of tiagabine HCl (TGB), a selective CNS GABA uptake inhibitor, in nearly 3100 patients from 53 separate clinical trials. TGB was found to have no clinically important effect upon hepatic metabolic processes, serum concentrations of concomitant antiepileptic drugs (AEDs), laboratory values, or important interactions with any common non-AEDs. Adverse effects were usually mild and involved the nervous system. TGB is safe and well-tolerated as add-on therapy for the treatment of partial seizures.     

Routine EEG vs. intensive monitoring in the evaluation of intractable epilepsy

Appropriate treatment of patients with intractable seizures requires precise identification of the type (or types) of seizure the patient experiences and correlation of this information with data from electroencephalography localizing the focus of the seizure in the brain. For such patients, the technique of "intensive monitoring" has gained rapid acceptance in the past several years as the investigative method of choice.Intensive monitoring usually entails prolonged electroencephalographic recording with simultaneous videotaping of the patient. Another common technique is prolonged monitoring of the patient's electroencephalogram (EEG) by radiotelemetry, during which time the patient is closely observed by trained personnel for suspected seizures.To compare the quality of information obtained from intensive monitoring with that from careful routine electroencephalography, the authors reviewed the medical records of 100 consecutive patients who had received both kinds of study after being referred for treatment in the special Epilepsy Treatment Unit of the University of Minnesota's Comprehensive Epilepsy Program (CEP).Success of each method was defined by ability to record an actual seizure. The routine EEG examination recorded actual seizures in 7 percent of patients in the study. With video EEG, following careful withdrawal of anticonvulsant drugs, seizures were recorded in 70 percent of patients. Telemetered EEG recorded seizure activity in 50 percent of those patients for whom the other two methods had failed to detect seizures.Intensive monitoring revealed that 60 percent of patients for whom the routine EEG study had recorded only one seizure type actually suffered from two or more types. Clinical diagnosis was changed in 84 percent of the patients. In this study, intensive monitoring was found to be far superior to the routine EEG examination as an aid to precise diagnosis of intractable seizure disorders.     

激素治疗增加绝经后健康妇女尿失禁的发病率和严重程度

问题：在绝经后健康妇女中，激素治疗（hormone therapy，HT）对尿失禁（urinary incontinence，UI）的效果如何？     

Extended‐release antiepileptic drugs: A comparison of pharmacokinetic parameters relative to original immediate‐release formulations

Many antiepileptic drugs (AEDs) have short half‐lives with large fluctuations in peak‐to‐trough plasma concentrations. Consequences of these pharmacokinetic (PK) properties may include adverse events (AEs) and breakthrough seizures, potentially leading to poor adherence. To address these challenges, newer formulations of these AEDs have been developed using unique extended‐release (ER) technologies. These technologies extend the dosing interval such that dosing frequency can be minimized, which may improve patient adherence. Available ER formulations have the potential to minimize the spikes in maximum plasma concentrations (C_max) at steady‐state that often contribute to AEs during treatment with immediate‐release (IR) products. In so doing, tolerability advantages may lead to increased AED effectiveness by improving adherence and allowing higher doses if clinically indicated. Direct PK comparison studies of IR and ER formulations (e.g., carbamazepine, divalproate sodium, lamotrigine, oxcarbazepine, levetiracetam, and phenytoin) have found that dose‐normalized ER formulations may or may not be bioequivalent to their IR counterparts, but most ER formulations have a lower fluctuation index ([C_max–C_min]/C_avg) compared with the IR versions. This results in flatter concentration‐time plots. Not all ER preparations improve the various PK parameters to the same extent, and PK nuances may impact the effectiveness, tolerability, and adherence rates of various ER formulations.     

Advances in antiepileptic drug treatments. A rational basis for selecting drugs for older patients with epilepsy

Incidence of epilepsy increases rapidly after age 65; recent studies indicate that approximately 10% of nursing home residents are being treated with antiepileptic drugs (AEDs). Almost all are being treated with first generation AEDs. The average nursing home patient receives six medications, has age-related changes in protein binding, decreases in hepatic and renal clearance, and may have alterations in gastrointestinal absorption. AEDs that do not have drug-drug interactions, are not metabolized by the liver, and are readily absorbed may offer benefits in this population. New studies are demonstrating that the first generation AEDs have a number of shortcomings for treating older patients, whereas some of the newer AEDs may overcome these limitations. This paper reviews the present knowledge base and compares properties of the first generation AEDs with newer agents to develop a more rational approach for drug selection in older adults.     

上皮样血管瘤16例临床与病理分析

 目的:探讨上皮样血管瘤（EH）的临床及组织病理学特征。 方法:回顾性分析16例已确诊患者的临床及病理资料,并对相关文献进行综述。 结果:16例患者中女8例,男8例;平均年龄（45.44±12.52）岁;病程3个月~20年;皮损主要表现为红色至暗红色丘疹、结节,可伴瘙痒和糜烂,好发于头皮、耳部,其中发生在头部10例、耳部5例、外阴1例;皮损单发者5例,多发者11例。皮损组织病理学检查均具有典型的血管增生,管壁增厚,内皮细胞呈上皮样突向管腔,管周大量淋巴细胞及嗜酸性粒细胞浸润。结论：上皮样血管瘤少见,诊断需要临床与病理密切结合。加强对该病的认识,可避免误诊误治。