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.     

Procarcinogen activation by rat and human mammary extracts

Sprague-Dawley rat mammary gland is extremely sensitive to tumorigenesisby single or multiple doses of several poly-cydk aromatic hydrocarbons.We obtained quantitative data on the in vitro mutagenic activationof several procarcinogens by 9000 g supernatant fraction (S9)from rat mammary gland using the Ames test. Mutagenic activationwas shown to be dependent on a nicotinamide adenine dinucleotidephosphate (NADPH) generating system. An S9 preparation frommammary tissue of lactating Sprague-Dawley rats was shown toactivate 2-aminoanthracene (2-AA). A polychlorinated biphenylmixture of 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin (TCDD) givento rats greatly raised the specific activity (rever-tant TA98colonies/mg S9 protein) of the mammary tissue using 2-AA asa test carcinogen, and permitted detection of 2, 4-diaminoanisole(DAA) and 2, 7-diaminofluorene (DAF) activation. Procarcinogens2-aminofluorine (2-AF), benzo[a]-pyrene (BP) and aflatoxin (AFL)Bl were not detectably activated by mammary gland. Mutagenesisproduced in mammary S9 activation of 2-AA, DAA or DAF was significantlyinhibited by alpha-naphthoflavone (NF) but was inhibited minimallyby metyrapone (MP). Human mammary tumor cell lines (734B, SkBr3,MDA-MD-330) possessed inducible procarcinogen metabolizing activitiessimilar to those found in S9 of rat mammary tissue. We demonstrateda simple and convenient use of the Ames test to characterizeactivation of many potential mutagens and carcinogens for mammarygland. When a test compound such as 2-AA was used, selectiveenzyme induction and inhibition was demonstrated.     

Laparoscopic modified taylor procedure in the treatment of duodenal ulcer: technique and outcome after 5-year follow-up

BACKGROUND: Recently, gastric stapling with posterior truncal vagotomy has been performed by laparoscopic surgery, as an alternative to highly selective vagotomy (HSV) and the Taylor procedure for the treatment of chronic duodenal ulcer. AIM: To investigate, after a mean 5-year follow-up, the effect of the stapling-modified laparoscopic Taylor procedure, on gastric secretion, emptying and reflux as well as clinical parameters and recurrence rates in patients treated for duodenal ulcer. METHODS: 16 patients, aged 38-66 years, were treated from January 1993 to January 1996 (median 60.5 months), by the laparoscopic stapling-modified Taylor procedure, using the Endo-GIA stapler device. Assessment of the results of gastric acid secretion, solid and liquid gastric emptying, enterogastric reflux, endoscopic findings and clinical parameters, using the Visick grading, was performed. RESULTS: Endoscopy found healing ulcer in 15 patients. One patient showed signs of chronic ulcerative disease without gastritis or pyloric stenosis indicative of progressive ulcerative diathesis and was classified as Visick III. 14 patients were classified as Visick I and 1 as II. The enterogastric reflux index ranged from 0 to 26%, basal and peak acid output were 1.4 +/- 0.6 and 11.7 +/- 6.1 mmol H(+)/h, respectively. The half-emptying times of the solid and liquid meal were 82 +/- 7 and 16 +/- 6 min, respectively. These results are likely to be similar to those obtained from series of patients who underwent HSV or Taylor procedure and are closed to those from healthy controls. CONCLUSIONS: The laparoscopic modified Taylor procedure, using the Endo-GIA stapler device allows a more rapid, technically easier and radical performance of the operation with excellent long- term results and should be included in the armamentarium of the treatment of chronic duodenal ulcer.     

Altered NMDA glutamate receptor antagonist response in recovering ethanol-dependent patients.

Ethanol is an antagonist of the N-methyl-D-aspartate (NMDA) glutamate receptor. Ethanol dependence upregulates NMDA receptors and contributes to crosstolerance with selective NMDA receptor antagonists in animals. This study evaluated whether recovering ethanol-dependent patients show evidence of a reduced level of response to the effects of the NMDA receptor antagonist, ketamine. In this double-blind study, 34 recently detoxified alcohol-dependent patients and 26 healthy comparison subjects completed 3 test days involving a 40-min infusion of saline, ketamine 0.1 mg/kg, or ketamine 0.5 mg/kg in a randomized order. Recovering ethanol-dependent patients showed reduced perceptual alterations, dysphoric mood, and impairments in executive cognitive functions during ketamine infusion relative to the healthy comparison group. No attenuation of ketamine-induced amnestic effects, euphoria, or activation was observed. The alterations in NMDA receptor function observed in recovering ethanol-dependent patients may have important implications for ethanol tolerance, ethanol dependence, and the treatment of alcoholism.     

Usefulness of thoracoscopic surgery in the diagnosis and management of thoracic diseases

BACKGROUND: Video-assisted thoracoscopic surgery (VATS) has been recently utilised in the diagnosis and management of thoracic diseases. In this article we report our series of patients with established indications for VATS treatment. METHODS: Over the past 6 years we performed 104 VATS procedures for diagnostic and therapeutic purposes in 95 men and 39 women. The specific indications for VATS were: lung biopsy for undiagnosed diffuse lung disease, mediastinal biopsy and cysts, pleural effusion, empyema, pneumothorax and bullous lung disease, pericardial effusion and cyst, parvertebral abscess and solitary pulmonary nodules. RESULTS: There was no operative mortality. Postoperative non-fatal complications were seen in 7 cases. The overall median duration of chest tube drainage was 2.5 days and the mean postoperative stay 3 days. In diffuse lung disease a tissue diagnosis was obtained in all cases. Definitive diagnosis in the patients with undiagnosed pleural effusion was obtained in 90% of cases and the overall diagnostic rate was 98.5%. The success rate of the empyema (stage II) treatment and the therapeutic procedures is 100% after a mean follow-up of 12 months (range 6-30). Conversion to thoracotomy was needed in 6 cases. In all patients the postoperative pain was controlled with intake of non-narcotic analgesics with satisfactory results. CONCLUSIONS: VATS is worth considering and has been established as procedure of choice, with exceptional results in various chest diseases such as undiagnosed pleural effusions, recurrent, post-traumatic or complicated spontaneous pneumothorax, stage II empyema, accurate staging for lung cancer in the resection of peripheral solitary pulmonary nodule less than 3 cm, and lung biopsy for pulmonary diffuse disease.     

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？     

The use of antidepressants in alcohol-dependent veterans

BACKGROUND: Although some research reports suggest antidepressants may be an effective treatment for alcohol dependence, their clinical use has been controversial. METHOD: By using comprehensive national administrative data from the Department of Veterans Affairs (VA) documenting psychiatric diagnoses and care under natural conditions in both mental health and non-mental health settings, the use of antidepressants was compared in alcoholic and non-alcoholic veterans. Data were collected from April 1, 2000, to Sept. 30, 2000. RESULTS: Overall, patients with alcohol dependence were significantly more likely to be prescribed antidepressants than individuals without alcoholism (38.9% vs. 31.2%), but multivariate analysis showed that this was attributable to the higher rate of comorbid psychiatric diagnoses in this group. After controlling for diagnosis and other potentially confounding factors, alcohol-dependent individuals treated in specialty mental health clinics were in fact less likely to be prescribed antidepressants (odds ratio = 0.95, p <.0001). Of patients treated with antidepressants, those with alcohol dependence were more likely to be prescribed newer antidepressants (odds ratio = 1.22, p <.0001). Among patients treated in non-mental health clinics, there were no significant differences in rate of antidepressant use between alcohol-dependent and non-alcohol-dependent individuals with mental illness after controlling for other factors. CONCLUSION: The principal finding is that a diagnosis of alcoholism, independent of other factors, is associated with a decreased likelihood of receiving antidepressant treatment in VA mental health clinics, suggesting that prescribers have not embraced reports of their efficacy in alcohol dependence. In fact, this may reflect a disinclination to use medications to treat alcoholism in specialty mental health clinics but not in non-mental health clinics.