Na+-依赖性谷氨酰胺载体ASCT2的生物学功能及表皮生长因子的调控作用

目的:Na -依赖性中性氨基酸载体ASCT2,是一种广谱的氨基酸载体,它的生理功能主要是转运肠腔内谷氨酰胺、丙氨酸、丝氨酸和半胱氨酸等中性氨基酸.本研究拟对ASCT2载体转运特性及表皮生长因子对其的调控机制作一探讨. 方法:体外培养Caco-2细胞,观察ASCT2对核素标记谷氨酰胺的转运特性、动力学特征及底物特异性;利用ASCT2多克隆抗体及TaqMan实时定量PCR方法,研究表皮生长因子对ASCT2功能的调控作用. 结果:Caco-2细胞中Na -依赖性3H-L-谷氨酰胺(50 μmol)的转运速率为(635±80.1) pmol/(mg protein·min),大部分中性氨基酸如丙氨酸、丝氨酸、半胱氨酸等可对其产生竞争性抑制;在表皮生长因子(100 μg/L)孵育下,可以提高其转运活性约100%(P＜0.01),Western-blot及real-time PCR显示其蛋白及mRNA表达增高. 结论:谷氨酰胺载体ASCT2对核素标记谷氨酰胺具有强大的转运活性,对中性氨基酸底物具有亲和特异性;表皮生长因子对其转运活性、蛋白表达、mRNA丰度具有上调作用.     

肠黏膜刷状缘中性氨基酸载体ASCT2的结构和功能特点

ASCT2是肠黏膜刷状缘的一种Na 依赖性中性氨基酸载体,它的功能主要是转运中性氨基酸,如丙氨酸、丝氨酸、半胱氨酸、苏氨酸、谷氨酰胺和天冬酰胺等.对ASCT2分子结构、功能特点和调控特性等的研究,有助于对肠道功能的认识,并为临床短肠综合征(SBS)及其他肠功能障碍疾病提供新的治疗对策.     

神经导向因子在缺血肠道及缺氧细胞中的表达及作用

目的:探讨神经导向因子(Netrin-1)在急性肠系膜缺血(AMI)病人肠道组织及低氧条件下人结直肠腺癌(Caco-2)细胞中的表达及其对肠道功能恢复和微血管新生的作用. 方法:采用免疫印迹、免疫组化技术分别检测46例肠系膜上静脉血栓病人缺血肠道Netrin-1表达情况,并分析其表达量与病人主要临床结果之间的关系;培养Caco-2细胞,应用免疫印迹、Real-time PCR和免疫荧光技术检测不同低氧时间下Caco-2细胞Netrin-1 mRNA和蛋白的表达. 结果:预后较好的病人肠道局部Netrin-1表达量明显高于有严重并发症(30 d内死亡、二次手术探查、全肠外营养(TPN)时间＜14 d、脓毒症、需机械通气的ARDS、MODS、SBS)的病人,差异有显著性统计学意义(P＜0.05).同时,Caco-2细胞低氧时间与细胞Nertin-1存在一定相关性,且随着低氧时间逐渐延长,Netrin-1 mRNA及其蛋白表达量逐渐增加. 结论:AMI病人肠道局部的Netrin-1可能参与微血管新生及血管网络的构建而保护已缺血肠道,进而减少肠坏死等并发症,促进肠道功能恢复.     

HIF-1、GLUT-1与子宫内膜癌的研究进展

缺氧诱导因子-1是在缺氧条件下广泛存在于哺乳动物和人体内的一种转录因子.缺氧诱导因子-1能启动靶基因转录,使机体适应缺氧的微环境,其还在维持氧稳定、细胞能量代谢、肿瘤血管生成及转移、细胞增殖与凋亡等多方面起着重要作用.葡萄糖转运蛋白-1是一种膜性易化葡萄糖转运蛋白,是已知的葡萄糖转运蛋白家族中分布最广的转运体.在缺氧缺血条件下,缺氧诱导因子-1a通过介导缺氧反应基因而上调葡萄糖转运蛋白-1的表达.葡萄糖转运蛋白-1和缺氧诱导因子-1a联合阳性表达可能成为临床恶性肿瘤诊断,特别是早期诊断的指标,联合检测这两种指标在反映癌症发生发展过程中起关键作用.该文就缺氧诱导因子-1、葡萄糖转运蛋白-1与子宫内膜癌发生发展关系的研究进展作以概述.     

肝细胞质膜上的谷氨酰胺载体

谷氨酰胺(Gln)是哺乳动物血浆中最丰富的氨基酸,并参与体内多种生理和病理过程.Gln必须首先进出功能细胞,才能发挥其功能,而此过程又必须依赖细胞膜上特异Gln载体的跨膜转运.肝是Gln代谢的主要器官,了解肝细胞质膜Gln载体的结构和功能特性,有助于深入理解Gln的代谢与功能.     

不同剂量左甲状腺素对缺氧缺血性脑损伤新生大鼠脑组织HIF-1α表达的调节

目的 探讨不同剂量左甲状腺素(L-T4)对缺氧缺血性脑损伤(hypoxia-ischemia brain damage, HIBD)新生大鼠脑组织缺氧诱导因子-1α(hypoxia-inducible factor 1α, HIF-1α)的调节作用。方法 7日龄SD大鼠随机分为假手术组、缺氧缺血组(HI)、溶剂组及L-T4低、高剂量组。低、高剂量L-T4干预组及溶剂对照组分别于HIBD后当日起腹腔注射2 μg/100 g L-T4、3.5 μg/100 g L-T4及等体积溶剂, 每日1次, 共5 d。应用免疫组化法检测鼠脑HIF-1α蛋白, RT-PCR法检测HIF-1αmRNA。结果 缺氧缺血组缺血侧大脑皮质HIF-1α蛋白(72.795±6.121)及HIF-1αmRNA(0.448±0.035)表达较假手术组(依次为38.581±2.846, 0.174±0.015)增高, 差异有统计学意义(P＜0.05);L-T4低、高剂量干预组缺血侧大脑皮质HIF-1α蛋白(依次为117.350±9.374, 142.842±8.948)及HIF-1αmRNA(依次为0.618±0.042, 0.711±0.049)表达较缺氧缺血组增高, 差异有统计学意义(P＜0.05), 且L-T4对HIF-1α的影响具有剂量依赖性, 即随剂量增加, HIF-1α蛋白及HIF-1αmRNA表达增加, 两组间差异具有统计学意义(P<0.05)。结论 在新生大鼠HIBD时, L-T4可上调HIF-1α蛋白及其mRNA表达, 从而对HIBD起保护作用, 且具有剂量依赖性。     

肠上皮细胞中性氨基酸载体B0AT1的真核表达及稳定转染体系的建立

目的: 构建可表达人中性氨基酸载体B0AT1基因的真核表达载体,建立重组质粒转染的Hela细胞系,筛选出稳定表达人B0AT1的细胞株. 方法: 提取人正常小肠上皮细胞总RNA,采用RT-PCR方法扩增出B0AT1基因片段,EcoRⅠ和XbaⅠ双酶切后,将其插入至真核表达载体pcDNA3.1中,构建重组表达质粒pcDNA3.1-B0AT1,转化E.coli DH5α菌株感受态细胞,将阳性克隆脂质体法转染Hela细胞,经G418筛选获得稳定表达株,用RT-PCR法检测B0AT1基因的mRNA表达. 结果: 从小肠上皮细胞中成功克隆到人B0AT1基因.酶切和序列测定表明已成功构建真核表达载体pcDNA3.1-B0AT1,将此重组质粒转染的Hela细胞,可稳定表达人B0AT1基因,并筛选出稳定表达B0AT1的Hela细胞系.氨基酸摄取实验证实,转染pcDNA3.1-B0AT1的Hela细胞具有B0AT1基因的生物学活性. 结论: 重组人中性氨基酸载体B0AT1克隆成功,并在Hela细胞中获得稳定表达.     

胎鼠宫内缺氧缺血后脑海马GAP-43 mRNA的表达

目的:探讨孕鼠子宫胎盘缺氧缺血对胎鼠脑海马发育的影响。方法:利用子宫动脉钳夹法对妊娠第17 d孕鼠实施单侧子宫动脉缺氧缺血手术,制成孕鼠子宫动脉缺氧缺血模型。缺氧缺血组孕鼠的另一侧子宫动脉不钳夹,相应为对照组。采用半定量RT-PCR方法分别检测两组胎鼠脑海马区GAP-43 mRNA,比较缺氧缺血组孕鼠子宫动脉钳夹缺氧缺血侧,与自身对照组孕鼠子宫动脉不钳夹侧的胎鼠海马脑组织生长相关蛋白-43(GAP-43)mRNA的表达。结果:缺氧缺血组与对照组胎鼠脑海马GAP-43 mRNA的表达分别为1.03±0.15和1.19±0.08,两组比较,差异有极显著性(P<0.01)。结论:妊娠期子宫内缺氧缺血使脑海马区GAP-43 mRNA的表达显著减少,提示妊娠期子宫内缺氧缺血对胎鼠脑海马的发育产生影响。     

缺氧对滋养细胞E-钙黏蛋白表达的影响

目的:了解在缺氧条件下,单层贴壁培养的滋养细胞系TEV-1中,E-钙黏蛋白(E-cadherin,E-cad)的表达在mRNA水平和蛋白水平变化情况,探讨缺氧环境对滋养细胞迁移能力的影响,深入了解缺氧环境诱发子痫前期发病的潜在分子机制。方法:体外贴壁培养的滋养细胞系(TEV-1),分别直接接种于24孔板进行细胞爬片和培养瓶内,分为常氧组和缺氧组。常氧组加新鲜DMEM/F12培养基,缺氧组加含终浓度为300μmol/L二氯化钴的培养基,然后在正常条件下(37℃,5%CO2)培养。两组细胞培养48 h后,分别收集细胞,采用Trizol法抽提mRNA,并进行RT-PCR,检测E-钙黏蛋白的mRNA表达水平;细胞爬片采用免疫组织化学法进行染色,检测E-钙黏蛋白的蛋白表达水平。统计数据分析两组差异性。结果:二氯化钴化学缺氧条件下培养后,滋养细胞E-钙黏蛋白的mRNA和蛋白表达水平均较常氧对照组升高,差异具有统计学意义(P<0.05)。结论:缺氧环境可能导致滋养细胞E-钙黏蛋白的mRNA和蛋白的表达水平升高,由此降低了滋养细胞的迁移能力。这可能是缺氧造成胎盘浅着床、继而发生子痫前期的重要分子机制。     

乳酸菌对感染肠上皮细胞通透性及紧密连接蛋白表达的影响

目的:研究乳酸菌对侵袭性大肠杆菌(EIEC) 感染肠上皮细胞(Caco-2)后,对其通透性和紧密连接(TJ)蛋白表达的影响. 方法:建立Caco-2 单层细胞感染模型.实验分为正常组、感染组、乳酸菌组和庆大霉素组.用电阻仪测定单层细胞的跨膜电阻值(TER),用高效液相法(HPLC)测定甘露醇透过率的变化,采用免疫组化法观察TJ相关蛋白,如Claudin-1蛋白,Occludin蛋白, JAM-1蛋白,ZO-1蛋白的分布和结构变化. 结果:EIEC感染Caco-2细胞后,单层细胞的TER值和甘露醇的透过率随时间延长而升高,而经乳酸菌处理后,TER值升高趋缓(P＜0.05),甘露醇的透过率降低(P＜0.05);EIEC感染后,相邻Caco-2细胞间TJ结构遭到破坏,TJ相关蛋白的表达亦减少,而乳酸菌处理后可使EIEC引起的TJ结构受损减轻、相关蛋白的表达增多. 结论:乳酸菌黏附于肠上皮细胞Caco-2后,可抑制EIEC破坏单层细胞的完整性,并改善TJ的结构变化和相关蛋白的表达分布.     

长春新碱过量引起严重毒副反应1例的护理体会

报道1例霍奇金淋巴瘤患者因误用长春新碱（VCR）10mg一次性静脉推注后治疗护理情况。其出现间断性神志恍惚、眼睑闭合不全、言语不清、口腔黏膜糜烂、全身疼痛、麻痹性肠梗阻、尿潴留、手足麻木等症状,经积极解救,禁食,持续胃肠减压、胃管内注入麻油、开塞露、生理盐水灌肠,合理应用肠外营养,注重疼痛、心理护理,做好口腔、肛周护理,预防感染加重,患者病情得到控制好转出院。     

Correlates of stages of change of smoking among inhabitants of deprived neighbourhoods

BACKGROUND: This study examines the prevalence and correlates of stages of change of smoking, in terms of psychosocial, structural and sociodemographic factors, among inhabitants of deprived neighbourhoods. METHODS: Cross-sectional data were obtained from a survey on health related behaviour. Subjects were 2009 current and former smokers, aged 20-46, living in deprived neighbourhoods in Rotterdam, the second largest city in the Netherlands. Three groups of smokers were formed according to the stages of change-definitions of the Transtheoretical Model: smokers not planning to quit (precontemplators), smokers planning to quit (contemplators/preparators) and former smokers (actors/maintainers). Smokers planning to quit and smokers not planning to quit were compared regarding psychosocial factors (attitude, social norm, self-efficacy), structural factors (neighbourhood problems, material deprivation, financial problems, employment status) and sociodemographic factors (age, gender, marital status, cultural background, educational level). Former smokers were compared with smokers planning to quit regarding structural and sociodemographic factors. Logistic regression was used to assess correlates of stages of change. RESULTS: Smokers planning to quit (prevalence = 19%) reported a more positive attitude, stronger social norms and higher self-efficacy expectations in quitting smoking than smokers not planning to quit (prevalence = 57%). Smokers planning to quit less often were Dutch-born, more often had attended higher vocational schooling or university and more often reported experiencing two or more neighbourhood problems compared to smokers not planning to quit. Former smokers (prevalence = 24%) were older, more often Dutch-born, married, employed and higher educated, compared to smokers planning to quit. Furthermore, former smokers less often reported material deprivation and financial problems than smokers planning to quit. CONCLUSION: Among people living in deprived neighbourhoods, different factors correlate with different stages of change of smoking. Implications for health promotion are discussed.     

Choice of measures of vaccination and estimates of risk of pediatric pertussis

BackgroundVaccination uptake at the individual level can be assessed in a variety of ways, including traditional measures of being up-to-date (UTD), measures of UTD that consider dose timing, like age-appropriate vaccination, and risk reduction from individual doses. This analysis compared methods of operationalizing vaccination uptake and corresponding risk of pertussis infection.MethodsCity-wide case-control study of children in Philadelphia aged 3 months through 6 years, between 2001 and 2013. Multiple logistic regression was used to isolate the independent effects of each measure of vaccination uptake and the corresponding relative odds of pertussis.ResultsBeing UTD on vaccinations was associated with a 52% reduction in risk of pertussis (OR 0.48, 95% CI: 0.34, 0.69). Evaluation of delayed receipt of vaccine versus on-time UTD yielded similar results. There was a decrease in risk of pertussis for each additional dose received with the greatest reduction in pertussis infection observed from the first (OR 0.48, 95% CI: 0.28, 0.83) and second dose (OR 0.17, 95% CI: 0.08, 0.34). Additional doses conferred minimal additional protection in this age group.ConclusionExamining vaccination status by individual doses may offer improved predictive capacity for identifying children at risk for pertussis infection compared to the traditional UTD measure.     

上海市某医院骨科平均住院日影响因素分析

对上海市某医院2003年-2007年骨科出院病人的住院日描述性分析．2003年-2007年骨科的床位利用指数与平均住院日相关性分析．2003年-2007年骨科床位与医护比例分析．2007年骨科前10大病种平均住院目影响因素分别进行单因素相关性分析和多因素逐步回归分析（STATA软件）。通过对骨科10大病种住院日影响因素分析,术前等待天数、手术类型、是否输血分别对10个、9个和8个病种的住院目有影响。输血因素和手术类型是医院不可控、由病人的病情决定的,术前等待天数是管理因素,是最值得医院重视的影响因素。     

两种不同肌瘤剔除术治疗子宫肌瘤的临床疗效比较

目的:比较腹腔镜子宫肌瘤剔除术与腹式子宫肌瘤剔除术的临床疗效。方法:回顾分析63例腹腔镜子宫肌瘤剔除术与61例腹式子宫肌瘤剔除术的临床资料,比较2种手术方式的手术时间、术中出血量、肛门排气时间、使用抗生素时间、术后感染率、体温恢复正常时间、住院时间等情况。结果:两组患者接受子宫肌瘤剔除术顺利完成,腹腔镜组手术时间长于开腹组(P0.05),腹腔镜组术中出血量及剔除肌瘤个数均少于开腹组(P0.05),术后腹腔镜组体温恢复正常时间、肛门排气时间、抗生素使用及住院时间均短于开腹组,两组患者最多随访6个月,腹腔镜组平均康复时间较开腹组短(P0.05),两组随访B超均未发现明显肌瘤结节。结论:腹腔镜子宫肌瘤剔除术损伤小,恢复快,住院时间短,是值得推广的微创手术。     

尘肺病患者145例死因调查

目的分析汉中市尘肺病的发生、发展和死亡原因，以提高尘肺病患者的生存质量。方法对145例尘肺病患者的死因进行回顾性分析，计算其平均发病年龄、死亡年龄及患尘肺病后的存活年限。结果矽肺的平均发病年龄和死亡年龄均早于煤工尘肺和石棉肺，尘肺病的主要死因为患肺部疾患，患尘肺病后的平均存活年限为12a。结论加强职业健康体检工作，对尘肺病人做到早发现早治疗和预防并发症的发生。     

性发育异常发病机制的研究进展

正常的性腺分化可分为3个过程：原始性腺形成、性别决定和卵巢/睾丸发育。在任一环节中,基因表达或调控发生异常均有可能导致性发育异常疾病的发生。性发育异常是指染色体、性腺和解剖性别不典型。随着分子生物学技术的发展,不断地发现新的基因或信号通路参与性腺分化和发育,如SRY、SF1、WT1、Sox9等基因与睾丸发育密切相关,Wnt/Rspo1/B连环蛋白通路、Dax1、Foxl2等基因在卵巢分化中发挥着重要作用,一些非编码RNA和转化生长因子也有重要的调节功能,且睾丸和卵巢发育均为主动过程,即使在出生后因某些基因的改变两者间也可出现横向分化。这些为揭示性分化异常的发病机制提供了可能。     

西林瓶装注射用水的装量检查分析

为了检查西林瓶装注射用水的实际分装量是否符合2005年版《药典》三部要求,按照2005年版《药典》三部中“生物制品分装和冻干规程”,分装规格为0．8ml的西林瓶装注射用水10000瓶,随机在分装过程的前、中、后阶段共抽取100瓶,轧盖、目检。分别用1ml标定注射器及1ml普通无菌注射器抽取每瓶注射用水的实际体积,以t检验法对实际抽取量与标示量的差值进行统计学分析,结果显示其与2005年版《药典》三部要求分装附加量为0．1ml相比有显著性差异（P〈0．001）。     

One-year stability of the Measure of Processes of Care

BACKGROUND: The Measure of Processes of Care (MPOC) is a 56-item self-administered measure designed to examine what parents of a child with a chronic health problem think of the services they and their child receive, and to measure the extent to which these services are family-centred. Reliability and validity of the MPOC were established in prior studies. The aim of the present study was to assess the 1-year stability of the MPOC to justify its use as an evaluative tool. METHODS: Nine paediatric rehabilitation centres in the Netherlands participated in this short longitudinal survey study. Subjects were 205 parents (response rate 74.8%) of children aged 1-18 years who received care in one of the participating paediatric rehabilitation centres. All subjects filled out two MPOCs with a 1-year interval. RESULTS: All correlations between the scale scores of the MPOC at the first and second administration were relatively high and significant (range: 0.443-0.609, all P < 0.001), demonstrating high inter-individual stability. However, all mean scale scores, except for Providing General Information, significantly reduced after 1 year. CONCLUSIONS: The MPOC has a moderate 1-year stability. However, because of its tendency to score lower when repeated after 1 year, its use as an evaluative follow-up instrument to assess the effectiveness of a programme intervention is restricted.     

儿童皮质发育畸形的MRI诊断

目的探讨儿童皮质发育畸形(MCD)的MRI表现特点,为MCD的早期诊断提供依据。 方法选择2010年1月至2012年12月在四川省医学科学院·四川省人民医院放射科经MRI检查确诊为MCD的44例患儿为研究对象。回顾性分析及总结其MRI影像学特征。本研究遵循的程序符合四川省医学科学院·四川省人民医院人体试验委员会制定的伦理学标准,得到该委员会批准,征得受试对象及其监护人知情同意,并与监护人签署临床研究知情同意书。 结果本组44例经MRI确诊为MCD患儿中,巨脑回-无脑回为17例,多小脑回为14例,灰质异位为9例,脑裂畸形为4例。MCD的MRI特点中,巨脑回表现为脑回增宽、皮质增厚及白质变薄;无脑回表现为正常脑沟、脑回消失,皮质增厚及白质变薄;多小脑回表现为脑回增多、细小;灰质异位表现为皮质下及侧脑室周围深部白质区结节状、团块状或带状病灶,信号与灰质信号相同;脑裂畸形表现为从侧脑室到脑表面贯穿大脑半球的裂隙,表面覆盖灰质。 结论MRI是诊断儿童MCD的可靠影像学方法。