口蹄疫病毒持续感染细胞系的快速选择及其特性研究

采用NH4Cl弱碱法处理感染口蹄疫病毒的叙利亚仓鼠肾细胞系(BHK-21), 存活细胞经过单克隆和选择,获得32株阳性克隆细胞株.随机选取一株(BHK-ROp),常规传代并采用RT-PCR,透射电子显微镜,流式细胞仪进行持续感染特性分析,结果表明,BHK-ROp第4,16,36代细胞中病毒持续存在,但不影响细胞的生长特性,表现出病毒持续感染的基本特征.可见,NH4Cl弱碱法用于建立病毒持续感染细胞系是十分有效的.     

人基质金属蛋白酶-2C端片段PEX的高效表达及活性研究

将人的PEX基因克隆到表达载体pET-15b上,构建重组工程菌BL21(DE3)/pETPEX。采用5L发酵罐补料分批培养,当菌体密度增长到OD₆₀₀为35时,用IPTG诱导,PEX蛋白形成包涵体,最终OD₆₀₀达到90,PEX的表达水平达2.2g/L。包涵体蛋白经过纯化、复性后,获得生物活性。PEX能够有效抑制HUVECs的增殖,加药96h后,20nmol/L PEX对HUVECs生长有77.9%的抑制率;PEX能阻断bFGF诱导的鸡胚尿囊膜血管生成,抑制率为85%;小鼠实验表明,当给药剂量为5mg/kg时,PEX对S180肉瘤有31.4%的抑瘤率(P<0.05)。PEX是一个有效的抑制新血管生成和肿瘤生长的蛋白质因子。     

Factors Affecting the Rate of Photophosphorylation by Isolated Chloroplasts of Euglena gracilis*

SYNOPSIS The rate of photophosphorylation by Euglena chloroplasts depends not only on the physiologic stage of cell growth but also on the stage of development of chloroplasts in these cells at the time of harvesting. Both of these processes can be markedly influenced by a number of environmental factors; they are affected neither in a parallel manner nor completely independent of each other. In addition, the rate of photophosphorylation of chloroplasts can also be greatly affected by the conditions employed for their isolation. After investigating the various environmental factors both during cell growth and chloroplast isolation, we have developed a procedure which increased the photophosphorylation rate of our chloroplast preparations more than 5-fold, giving a specific activity in the range of 100-150 μmoles ATP/mg chlorophyll/hr routinely. The procedure is simple, needs no special equipment and requires only 2 or 3 days for cell growth.     

Regulation of Crustacean Molting: A Multi-Hormonal System

SYNOPSIS. In order to increase in size, arthropods must firstmolt (shed) their confining exoskeleton. This molting processis under the immediate control of the steroid molting hormone20-hydroxyecdysone (20-HE). Both the initial rise in circulatinghormone concentration and a coordinated decline are necessaryfor successful molting. Synthesis and/or release of ecdysone,the precursor to 20-HE, is regulated by the neuropeptide molt-inhibitinghormone (MIH). We have determined the primary amino acid sequenceof MIH in the lobster, Homarus americanus. This peptide hasa high degree of identity with the lobster hyperglycemic hormone.Another endocrine factor that appears to be involved in moltingis the juvenile hormone-like terpenoid methyl farnesoate (MF).We have characterized hemolymph MF binding proteins during themolt cycle. In addition, recent data indicate that MF may stimulatethe secretion of 20-HE in vivo and in vitro.     

Amino-acid Sequence of Substance P

IN 1931 von Euler and Gaddum¹, studying the tissue distribution of acetylcholine, found that brain and intestine contained a substance that stimulated contraction of the isolated rabbit jejunum and caused transient hypotension when injected intravenously into anaesthetized rabbits. These effects could not be ascribed to acetylcholine, for they were not prevented by the previous administration of atropine. The initial studies were made on crude acid alcohol extracts of equine brain and intestine, dried in powder form. The active principle in the preparation was later referred to as substance P (P for powder) and this non-committal term subsequently achieved widespread acceptance in the literature, in the absence of any clearly definable biological role for the compound (or compounds) involved.     

拟南芥高丝氨酸激酶HSK的分子结构与同源建模研究

高丝氨酸激酶（EC2．7．1．39）是拟南芥中合成甲硫氨酸和苏氨酸的关键酶之一。作者利用DNAMAN软件分析高丝氨酸激酶的氨基酸组成和等电点;利用生物信息学在线网站预测分析其疏水性、二级结构、跨膜结构域、信号肽区域、PEST序列、化学修饰位点及亚细胞定位等;利用SWISS—MODEL软件进行同源建模,使用Chimera（V1．8．1）做结构修饰处理,最后利用Molprobity4及ERRAT（V2．0）对建模结果进行结构质量分析。结果发现：HSK预测等电点为8．48,平均亲水性0．267;二级结构中α-螺旋占34．32％,不规则卷曲占48．11％,延伸链占17．57％;高丝氨酸激酶的预测模型经验证具有稳定的空间结构,符合立体化学规则。     

青年妇女患宫颈癌的临床特点及预后分析

目的：探讨青年妇女患宫颈癌的临床特点并行预后分析。方法：选取2008年1月-2013年1月在我院接受治疗的青年妇女宫颈癌33例患者观察组,同期在我院接受治疗的大于35岁49例患者为对照组。对两组患者的发病相关因素及临床病理特征进行回顾性分析及对两组患者预后分析,随访患者5年内生存率及2年内复发率。结果：两组患者初次性行为差异无统计学意义（P〉0．05）;两组患者避孕方式与妊娠次数比较均有显著性差异（P〈0．05）;两组患者临床分期与宫颈形态无显著性差异（P〉0．05）;两组癌灶大小、病理分型、宫颈肌层浸润深度及盆腔淋巴转移具有显著性差异（P〈0．05）;观察组较对照组患者早期（Ⅰa—Ⅱa）与晚期（≥Ⅱb）的两年内复发率及五年生存率差异均有显著性差异（P〈0．05）。结论：青年妇女宫颈癌患者具有癌灶大、腺癌发病率高、宫颈肌层浸润深度≥1／2肌层和盆腔淋巴结转移阳性的临床特点。     

小剂量右美托咪定用于局麻下玻璃体切割术的镇静效果观察

目的：探讨右美托咪定用于局麻下玻璃体切割术的镇静效果。方法：选择拟在局麻监测下行玻璃体切割术患者50 例为研究对象,年龄20-72岁,ASA 分级Ⅱ级~Ⅲ级,随机分为右美托咪定组（D组）和咪达唑仑组（M 组）,每组25 例。D组患者于术前10min 静脉泵注右美托咪定0.5 滋g/kg,后以0.2-0.4 μg/kg.h 的速度持续输注,M组术前10 min 缓慢静脉注射咪达唑仑0.02 mg/kg,术中按需静注0.5 mg/ 次。维持VAS 评分≤ 4 分,Ramsay评分2-4 分。记录和比较两组患者术中血压、心率、呼吸的变化、辅助用药及患者对镇静效果的满意度。结果：给药后,M组T5时点MAP较T0显著下降（P〈0.05）,D组T10及以后各时点MAP 较T0显著下降（P〈0.05）;D 组T5及以后各时点BP 较T0显著下降（P〈0.05）,但组间及M 组组内BP 比较差异无统计学意义（P〉0.05）;M 组T30时点HR 较T0 显著下降（P〈0.01）,而在T5时,组间比较差异有统计学意义（P〈0.01）,即D 组下降更为显著。给药后各时点,D 组VAS 评分均显著低于M组（P〈0.05）,30 min 时达最低。两组Ramsay镇静评分给药后5 min 均达2 级以上,与给药前比较均显著升高（P〈0.05）,D组给药后30 min 及以后各时点Ramsay镇静评分均显著高于M组（P〈0.01）。给药后各时点,两组组内和组间SPO2和RR 比较均无统计学差异（P〉0.05）。D 组患者满意度较M组更高（P〈0.05）。结论：小剂量右美托咪定用于玻璃体切割术可使患者血流动力学平稳,镇静效果良好,疼痛感觉减轻,舒适度提高。     

腹腔镜手术治疗肾盂旁囊肿的体会(附67例报告)

目的:探讨腹腔镜技术治疗肾盂旁囊肿的方法和疗效,并结合治疗体会,为临床提供更多的参考依据。方法:对2006年3月~2011年9月我科收治的67例肾盂旁囊肿患者的临床资料进行回顾性分析,其中采用经腹膜后途径29例,经腹腔途径20例,单孔腹腔镜技术18例。分析患者的手术时间、术中出血量、并发症的发生情况、术后住院时间及复查随访情况。结果:所有病例的手术均成功,手术时间38~120min,平均时间(70.3±9)min;术中出血量10~125 ml,平均(47.3±15.7)ml;均未出现肾蒂及肾盂损伤等并发症。术后2~6天(平均3.4天)拔除引流管,术后住院3~7 d,平均(4.2±0.6)d。术后1月复查,腰酸、腰痛、镜下血尿以及肾积水、高血压等临床症状均消失。术后随访1~47个月,未见囊肿复发。结论:腹腔镜手术治疗肾盂旁囊肿具有暴露充分、创伤小、出血少、恢复快等优点,可作为肾盂旁囊肿治疗的首选方法。术者应严格掌握其适应症,术前完善相关检查,术中仔细辨认肾静脉壁与囊肿,操作轻柔,填入肾门脂肪可有效防止囊肿的术后复发。