microRNA-19对肝癌细胞系迁移与侵袭能力的影响

目的 比较不同转移潜能肝癌细胞系中microRNA-19（mir-19）表达变化,探讨mir-19对不同转移潜能肝癌细胞迁移和侵袭能力的影响.方法 应用TaqMan MGB探针法定量分析mir-19在高转移潜能肝癌细胞系（MHCC97H、LM3）和低转移潜能肝癌细胞系（HepG2、7402）的表达差异.利用转染试剂将人工合成的mir-19的前体转染高转移潜能细胞系,人工合成的mir-19抑制剂转染低转移潜能细胞系.采用细胞计数试剂盒-8（Cell Counting Kit-8,CCK-8）检测mir-19对细胞增殖的影响,Transwell小室检测mir-19对细胞的迁移和侵袭能力的影响.结果 在不同转移潜能的细胞系中,mir-19表达水平在高转移潜能细胞系较低转移潜能细胞系表达下调（P〈0.05）.功能实验显示mir-19可抑制肝癌细胞的侵袭和增殖（P〈0.05）,利用生物信息学预测mir-19的靶基因发现,许多癌基因3＇UTR包含与mir-19的互补序列.结论 mir-19可能部分通过调节CREB3L4、PDE4A、JAM2、RUNX2等的表达来抑制肝癌细胞的增殖和侵袭转移.     

GATA4在肝癌细胞中的表达及其对肝癌细胞增殖、迁移和侵袭能力的影响

目的探索转录因子GATA4在肝癌细胞系中的表达及其表达改变对肝癌细胞增殖、迁移和侵袭能力的影响。方法通过RT-PCR和Western Blot检测正常肝细胞、肝癌细胞系的表达,在体外构建GATA4基因沉默和过表达模型,分析并比较转录因子GATA4表达变化对肝癌细胞增殖、迁移和侵袭的影响。结果 GATA4mRNA在肝癌细胞系(SMMC7721、Hep3B)中高表达,在正常肝细胞HLLP-7702和HepG2中低表达,GATA4蛋白在正常肝细胞(HL-7702)和肝癌细胞系(SMMC7721、Hep3B和HepG2)中均低表达;GATA4高表达抑制肝癌细胞(HepG2)增殖、迁移和侵袭(P0.05),GATA低表达促进肝癌细胞(Hep3B)增殖、迁移和侵袭(P0.05)。结论 GATA4mRNA在部分肝癌细胞系中高表达,GATA4蛋白质水平在肝癌细胞系和正常肝细胞中均低表达。上调GATA4表达水平可显著抑制肝癌细胞(HepG2)增殖、迁移和侵袭能力,相反,下调GATA4的表达水平增强肝癌细胞(Hep3B)的增殖、迁移和侵袭能力,确切机制有待进一步探讨。     

OTUB2表达下调对胃癌细胞增殖、迁移、侵袭能力及PI3K/AKT信号通路的影响

目的探讨含OTU结构域的泛素醛结合蛋白2(OTUB2)表达下调对胃癌细胞增殖、迁移、侵袭能力及磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(AKT)信号通路的影响。方法采用Lipofectmine 2000转染试剂向MGC-803细胞中转染OTUB2小干扰RNA(si-OTUB2)序列(si-OTUB2组)和si-NC序列(si-NC组),以不进行任何转染的胃癌MGC-803细胞作为空白对照组。分别采用实时荧光定量PCR和蛋白质印迹法(Western blot)检测细胞中OTUB2mRNA及蛋白的表达水平。采用CCK-8法检测细胞增殖能力,Transwell小室检测细胞迁移和侵袭能力,Western blot检测增殖细胞核抗原(PCNA)、E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、AKT、磷酸化蛋白激酶B(p-AKT)蛋白表达水平。结果与空白对照组和si-NC组比较,siOTUB2组细胞中OTUB2mRNA及蛋白表达水平降低(P0.05),细胞增殖能力减弱(P0.05),细胞迁移数和侵袭数减少(P0.05),细胞中PCNA、N-cadherin和p-AKT蛋白表达水平降低(P0.05),E-cadherin蛋白表达水平升高(P0.05)。结论下调OTUB2可以抑制胃癌细胞的增殖、迁移、侵袭,其作用机制可能与抑制PI3K/AKT信号通路有关。     

过表达HIF-2α对肝癌细胞运动和侵袭的影响

目的:探讨缺氧诱导因子(hypoxia inducible factor-2α,HIF-2α)对肝癌细胞运动和侵袭的影响。方法:构建HIF-2α过表达载体,转染肝癌细胞,构建过表达稳定表达细胞株。采用实时荧光定量多聚酶联反应(quantitative real-time polymerase chain reaction,qRT-PCR)和蛋白质印迹法检测HIF-2α表达水平,通过Transwell运动实验、划痕实验体外分析过表达HIF-2α对肝癌细胞运动的影响;将过表达HIF-2α细胞原位接种裸鼠,观察小鼠肺脏的转移情况。采用蛋白质印迹法检测EMT相关指标分析HIF-2α促进肝癌细胞运动、侵袭能力的作用机制。结果:将过表达载体转染肝癌细胞后,HIF-2αmRNA和蛋白表达水平均升高(P0.05)。Transwell运动实验发现,高表达HIF-2α后,穿膜细胞数目明显增加(P0.01);划痕实验结果也表明高表达HIF-2α的细胞运动能力增强。体内侵袭实验发现高表达HIF-2α组小鼠肺转移灶大小增加。蛋白质印迹结果显示HIF-2α可促进间质细胞相关标志物蛋白表达增加。结论:过表达HIF-2α可能通过促进肝癌细胞的EMT转化提高肝癌细胞的运动、侵袭能力。     

YB-1对卵巢癌细胞EMT标志蛋白表达及黏附能力的影响研究

目的研究Y-box结合蛋白(YB-1)对卵巢癌细胞上皮间质转化(EMT)标志蛋白表达及细胞黏附能力的影响。方法在卵巢癌细胞中转染p Genesil-1.1/YB-1 shRNA和p Genesil-1.1/shRNA control,筛选稳定转染的细胞记为YB-1 shRNA组和shRNA-NC组,把没有转染的细胞作为对照组。以Realtime PCR和Western blot方法检测细胞中YB-1表达水平。用四甲基偶氮唑盐比色法(MTT)方法分别检测细胞增殖和黏附能力,用Western blot检测EMT标志蛋白上皮钙黏附素(E-cadherin)、神经型钙粘素(N-cadherin)和迁移侵袭相关蛋白基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)水平,Transwell小室测定细胞侵袭和迁移情况。结果 YB-1 shRNA组细胞中的YB-1表达水平明显低于对照组和shRNA-NC组(P0.05)。YB-1 shRNA组细胞增殖率和黏附率低于对照组和shRNA-NC组(P0.05)。YB-1 shRNA组细胞中E-cadherin蛋白水平高于对照组和shRNA-NC组,N-cadherin、MMP-2、MMP-9蛋白水平低于对照组和shRNA-NC组,差异具有统计学意义(P0.05)。YB-1 shRNA组细胞侵袭和迁移数目低于对照组和shRNA-NC组(P0.05)。结论下调YB-1促进卵巢癌细胞E-cadherin表达,抑制N-cadherin表达,抑制细胞EMT,降低细胞黏附、侵袭、迁移能力。     

MTA1基因沉默对宫颈癌细胞增殖及转移能力的影响

【】 目的 探讨MTA1基因沉默对宫颈癌细胞转移增殖的影响。方法 用慢病毒转染法稳定转染Siha细胞,转染同时进行实验分组。用RT-PCR技术和蛋白印迹法测定宫颈癌Siha、Hela细胞中MTA1 mRNA和蛋白的表达。Transwell体外侵袭实验检测转染后Siha细胞的迁移能力,MTT检测法和克隆形成实验法检测转染后Siha细胞的细胞增殖能力,FACS细胞凋亡法检测转染后Siha细胞的细胞凋亡率,PI-FACS细胞周期法检测转染后Siha细胞的各个生长周期的细胞数。结果 (1)Transwell体外侵袭实验：相比NC组,KD组Transwell转移率经T-Test分析P=2.61E-08<0.05。(2)MTT检测结果表明：相比NC组,KD组细胞增殖减缓。(3)克隆形成实验结果显示：相比NC组,KD组克隆数经T-Test分析P值=0.0006<0.05。(4)FACS细胞凋亡：相比两个对照组,KD组凋亡率经T-Test分析P<0.05。结论 MTA1基因促进宫颈癌细胞的转移和增殖,沉默MTA1基因表达能使宫颈癌细胞增殖及迁移能力下降,加速宫颈癌细胞的凋亡,为抑制肿瘤转移奠定实验基础,最后为宫颈癌的新型药物性靶向治疗提供有力的实验依据。     

siRNA干扰技术沉默p66SHC对结直肠癌细胞生物学特性的影响

目的探讨干扰p66shc基因表达对结直肠癌HCT116细胞增殖、迁移、迁移的影响。方法实验分为对照组(不做任何处理)、NC-siRNA组和p66SHC-siRNA组,NC-siRNA组、p66SHC-siRNA组通过Lipofectamine 2000将特异性siRNA转染入结直肠癌HCT116细胞中。荧光定量PCR(RT-PCR)和免疫印迹试验(Western Blot)检测细胞中p66SHC的表达量;四甲基偶氮唑(MTT)法检测细胞的增殖能力;划痕实验和Transwell法观察细胞迁移、侵袭的变化;Western Blot检测细胞EMT相关蛋白钙黏蛋白E(E-cadherin)、波形蛋白(Vimentin)、钙黏蛋白N(N-cadherin)的水平。结果采用siRNA干扰HCT116细胞中p66SHC蛋白的表达后,细胞的增殖能力显著下降(P0.05),明显抑制细胞侵袭、迁移能力(P0.05),E-cadherin蛋白的水平明显上调(P0.05),Vimentin、N-cadherin蛋白的表达水平显著下调(P0.05)。结论干扰p66SHC基因的表达能阻碍EMT的发生,从而降低结直肠癌细胞的增殖、侵袭以及迁移能力。     

过表达FOXB2对肝癌细胞的增殖、迁移和侵袭的影响

目的 探讨叉头框蛋白B2（FOXB2）对肝癌细胞增殖、迁移和侵袭能力的影响。方法 采用定量PCR检测正常肝细胞LO2细胞和肝癌细胞系Hep3B、Huh7细胞FOXB2 mRNA的表达,构建过表达FOXB2的细胞模型,逆转录实时荧光定量PCR和蛋白免疫印迹法检测过表达效率,细胞计数试剂盒-8（CCK-8）法检测FOXB2对Huh7细胞增殖能力的影响;平板克隆实验检测FOXB2对Huh7细胞克隆形成能力的影响;Transwell实验检测FOXB2对Huh7细胞迁移和侵袭能力的影响。结果 FOXB2在肝癌细胞系Huh7和Hep3B细胞中的表达水平低于正常肝细胞LO2（P<0.001）;过表达FOXB2抑制了Huh7细胞的增殖（P=0.005）和克隆形成（P<0.001）;FOXB2过表达肝癌细胞的迁移（P<0.001）和侵袭（P=0.002）能力低于Vector对照细胞。结论 FOXB2在肝癌细胞中低表达,FOXB2过表达抑制了肝癌细胞的增殖、迁移和侵袭。     

长链非编码RNA LINC00473对人胃癌细胞增殖和迁移能力的影响

目的探讨LINC00473在人胃癌细胞系中的表达水平及其对胃癌细胞生物学行为的影响。方法采用实时荧光定量PCR检测胃癌细胞中LINC00473的表达水平;转染靶向LINC00473的siRNA片段或LINC00473过表达载体以构建敲减或过表达的胃癌细胞系;通过CCK-8细胞增殖实验、平板克隆形成实验、Transwell迁移实验、western blot实验检测细胞的增殖和迁移能力以及EMT相关蛋白标志物表达水平的变化。结果与GES-1细胞相比,LINC00473在胃癌细胞中的表达水平明显降低(P0.05)。与siNC转染对照组相比,敲减LINC00473表达后,靶细胞的增殖能力(F=163.10,P0.01)、克隆形成(t=3.29,P0.05)和迁移能力(t=4.68,P0.05)明显增加;E-cadherin表达减弱(t=4.08,P0.05),N-cadherin(t=5.06,P0.01)、Snail(t=7.69,P0.01)和Vimentin(t=3.82,P0.05)蛋白的表达水平升高。与Vector转染对照组相比,上调LINC00473表达后靶细胞的增殖能力(F=186.00,P0.01)、克隆形成(t=3.22,P0.05)和迁移能力(t=5.52,P0.01)明显下降;E-cadherin表达升高(t=2.90,P0.05),N-cadherin(t=7.44,P0.01)、Snail(t=2.78,P0.05)和Vimentin(t=4.64,P0.01)蛋白的表达水平降低。结论敲减LINC00473的表达可促进胃癌细胞的增殖、迁移的能力,可能通过调控EMT参与胃癌细胞的迁移。     

MTA1基因沉默对宫颈癌细胞增殖及转移能力的影响

目的:探讨MTA1基因沉默对宫颈癌细胞转移增殖的影响。方法:用慢病毒转染法稳定转染Siha细胞,转染同时进行实验分组。用RT-PCR技术和蛋白印迹法测定宫颈癌Siha、Hela细胞中MTA1 m RNA和蛋白的表达。Transwell体外侵袭实验检测转染后Siha细胞的迁移能力,MTT检测法和克隆形成实验法检测转染后Siha细胞的细胞增殖能力,FACS细胞凋亡法检测转染后Siha细胞的细胞凋亡率,PI-FACS细胞周期法检测转染后Siha细胞的各个生长周期的细胞数。结果:(1)Transwell体外侵袭实验:相比NC组,KD组Transwell转移率经T-Test分析P=2.61E-080.05。(2)MTT检测结果表明:相比NC组,KD组细胞增殖减缓。(3)克隆形成实验结果显示:相比NC组,KD组克隆数经T-Test分析P值=0.000 60.05。(4)FACS细胞凋亡:相比两个对照组,KD组凋亡率经T-Test分析P0.05。结论:MTA1基因促进宫颈癌细胞的转移和增殖,沉默MTA1基因表达能使宫颈癌细胞增殖及迁移能力下降,加速宫颈癌细胞的凋亡,为抑制肿瘤转移奠定实验基础,最后为宫颈癌的新型药物性靶向治疗提供有力的实验依据。     

现代残疾康复理念、政策与社区康复体系研究

本研究分析了当代残疾康复的理念和理论架构,构建了以国际公约、决议和政策、国内法律和政策以及操作性工具3个层次的康复理论体系;运用包容性发展,分析探讨了当代社区康复的体系和特点;就发展康复事业、为残疾人提供全面系统的康复服务提出相关的政策建议。     

国内外唇腭裂专科护理发展现状的文献研究及分析

目的了解和分析国内唇腭裂专科护理工作发展的现状。方法检索并分析万方医学网、中国知网、Springer Link及PubMed英文数据库近6年有关唇腭裂手术的围术期护理、患儿的喂养、手术安全管理、围术期疼痛研究以及心理护理的相关文献。结果共检索出国内文献89篇(核心期刊20篇)、国外文献22篇,其中围术期护理38篇、患儿的喂养16篇、手术安全管理10篇、围术期疼痛研究10篇、心理护理15篇。结论与国外文献相比,国内论文总结和回顾性居多,应增加研究探讨性论文,从而促进唇腭裂临床护理工作专科化的发展。     

城乡学童个性行为特征调查

目的探讨农村与城市儿童的个性行为特征。方法将97名农村学龄儿童设为农村组,97名城市学龄儿童设为城市组,采用艾森克个性问卷一儿童版和Achenbach’s 行为量表一儿童版进行评定分析。结果两组学龄儿童艾森克个性问卷各维度评分均无显著性差异（P均〉0．05）,且与全国常模相一致（P均〉0．05）;农村组男学童Achenbach’s行为量表违纪行为、攻击行为及外向型因子分均显著高于城市组,女学童退缩、社交问题因子分均显著高于城市组（P〈0．05或0．01）,其他因子分均无显著性差异（P均〉0．05）。结论农村与城市儿童个性无显著差异,而农村儿童存在较多的行为问题,可能与其所受教养和生活方式有关。     

Comparison of short-term and long-term protocols for stabilization and preservation of RNA and DNA of Leishmania, Trypanosoma, and Plasmodium

Molecular tools continue to be important in the prevention and control of parasitic diseases. However, using these techniques directly in the field remains a major challenge. Therefore, the preservation of clinical samples collected from endemic field areas for later analysis remains an important preanalytical process. This study aimed at identifying a suitable protocol for stabilization and preservation of RNA and DNA in bioclinical specimens for Trypanosoma, Leishmania, and Plasmodium research. Both spiked and unspiked blood samples were preserved in 7 protocols (different media; storage temperatures). Samples were evaluated for possible degradation of DNA and RNA along the storage duration up to the 10th week. Nucleic acid targets were assessed as follows: (i) Trypanosoma and Plasmodium RNA analysis was done using real-time nucleic acid sequence-based amplification (RT-NASBA) for 18S rRNA and for stage-specific Pfs25 mRNA, respectively; (ii) Trypanosoma DNA assessment analysis was conducted by using a conventional PCR for 18S rDNA; (iii) Leishmania RNA analysis was performed with a quantitative NASBA for 18S rRNA and Leishmania DNA assessment with an RT-PCR for 18S rDNA. Findings suggested that a newly developed L3™ buffer proved to be reliable and suitable for both short- and long-term preservation of parasite nucleic acid material. This buffer is envisaged to be suitable for utilization in field situations where resources are limited.     

Rates and risks of transmission of smallpox and mechanisms of prevention

In 1980, the World Health Organization declared smallpox eradicated from the world; the last known natural case had occurred in Somalia in 1977, and the United States had stopped routinely vaccinating its citizens in 1972. However, with increasing concerns regarding domestic and international terrorism, smallpox has resurfaced as a potential threat to global health. We review the direct and indirect modes of smallpox transmission and how patterns of transmission vary substantially, depending on the severity of circulating disease, vaccination status, environmental and socioeconomic factors, and the setting of an outbreak. We examine mechanisms for controlling outbreaks of disease and preventing further transmission in the event of an outbreak, with an emphasis on smallpox vaccination.     

Prevalence and severity of nausea and vomiting of pregnancy and effect of vitamin supplementation.

OBJECTIVE: Although nausea and vomiting of pregnancy is the most common medical condition during pregnancy, there are many unanswered questions regarding its cause, epidemiologic features and optimal management. The objectives of this study were to ascertain the prevalence of nausea and vomiting in a sample of Canadian women, to characterize the distribution of their severity and to investigate the role of vitamin B6 deficiency in their etiology. DESIGN: Prospective study. SETTING: Antenatal counselling service for pregnant women. PATIENTS: Three cohorts of women: a prospective, population-based cohort of 193 women, to estimate the rate and severity of nausea and vomiting (cohort A); a cohort of 555 women who sought advice for nausea with or without vomiting, to study the correlation between the maximal daily number of episodes of vomiting and maximal weight loss (cohort B); and a prospective cohort of 301 women who reported vomiting, to correlate vitamin supplementation with vomiting (cohort C). INTERVENTIONS: All 3 cohorts were interviewed during the counselling session, and cohort B was followed up prospectively. OUTCOME MEASURES: Frequency of nausea and vomiting, weight loss, maximal number of daily episodes of vomiting, rate of multivitamin supplementation. RESULTS: Overall, 67% of the women in cohort A reported experiencing nausea or vomiting, or both; 22% reported vomiting, and 9% experienced weight loss. In cohort B there was a significant correlation between the maximal number of daily episodes of vomiting and maximal weight loss, although there was wide variation (r2 = 0.25, p < 0.001). There was a highly significant correlation between the number of daily vomiting episodes and mean weight loss (r2 = 0.99). In cohort C, vomiting was significantly associated with lack of supplementation with multivitamins before 6 weeks' gestation (p = 0.002). CONCLUSIONS: The relation between number of daily vomiting episodes and mean weight loss may serve as a clinical tool to assess the severity of nausea and vomiting in pregnancy and the success of anti-emetics and rehydration regimens. Further study is needed to elucidate the biologic basis of the observed association between vomiting and lack of multivitamin supplementation in early pregnancy.     

History of violence and subjective health of mother and child

Objective: To study the self-reported prevalence of experienced violence among a cohort of women about two years after giving birth, their health during pregnancy, pregnancy outcomes and their experience of their child’s health.

Setting and subjects: In 2011, a total of 657 women participated in phase III of the Childbirth and Health Cohort Study in Icelandic Primary Health Care, 18 to 24 months after delivery. The women had previously participated in phase I around pregnancy week 16 and phase II 5–6 months after delivery. Data were collected by postal questionnaires.

Main outcome measures: Women’s reported history of experienced violence, sociodemographic and obstetric background, self-perceived health, the use of medications and their child’s perceived health.

Results: In phase III, 16% of women reported experiencing violence. These women felt less support from their current partner (p?p?p?p?p?p?=?0.008).

Conclusions: Our study confirms that a history of violence is common among women. A history of violence is associated with various maternal health problems during and after pregnancy, a higher rate of caesarean sections and maternal reports of health problems in their child 18–24 months after birth.

• KEY POINTS

• Violence is a major concern worldwide. Understanding the impact of violence on human health and developing effective preventive measures are important elements of any public health agenda.

• ??The reported prevalence of experiencing violence was 16% among women attending antenatal care in the primary health care setting in Iceland.

• ??Women with a history of violence reported worse health in general during pregnancy and delivered more often by caesarean section, compared to women with no such history.

• ??Mothers with a history of violence also evaluated the general health of their child as worse than women with no such history.

• ??The findings of this study support the importance of recognizing and addressing experienced violence among women in primary care.

     

我院文职护士管理和培训的实践与体会

目的构建文职护士培训、使用及管理的科学方法。方法通过规范管理、科学施训、作为骨干大胆任用等多种管理方式,使文职护士得到充分的发展。结果文职护士群体成长为医院优秀骨干,首批满3年的文职护士顺利通过续聘考核。结论科学的管理培训是文职护士成长的重要基石,对医院护理管理有着重要作用。     

通用设计原则与产品研发策略

本文介绍了通用设计的概念和设计原则,并阐述了根据此原则实施通用设计的产品研发策略。通用设计惠及的不仅是残疾人,也惠及了包括普通公众在内的广泛人群,以及企业。应将通用设计的原理应融入政府相关政策和未来发展的规划中,为残疾人以及更多人群提供合理便利的通用设计产品。     

骨脂肪瘤及血管脂肪瘤的CT及MRI表现:9例报告并文献复习

目的探讨骨脂肪瘤及血管脂肪瘤的CT及MRI表现。方法收集手术病理证实的9例骨脂肪瘤及血管脂肪瘤的临床及影像学资料,9例术前8例接受CT扫描,3例接受MRI检查。结果9例中骨脂肪瘤6例,骨血管脂肪瘤3例。6例骨脂肪瘤发生于骨内4例,骨旁2例。骨内脂肪瘤跟骨1例,距骨1例,眶骨1例,肱骨1例。骨旁脂肪瘤颅骨1例,股骨1例。骨血管脂肪瘤发生于肋骨1例,颅骨1例,椎骨1例。骨内脂肪瘤CT显示病变骨质呈轻度膨胀性改变,边缘见轻度硬化环,病灶内见脂肪密度及条弧形、结节状钙化,MRI显示病灶T1WI呈高信号,PDWI及T2WI抑脂序列呈低信号。骨旁脂肪瘤CT呈不均匀高密度团块,MRI显示病灶T1WI呈高信号,T1WI及T2WI抑脂序列呈低信号。3例骨血管脂肪瘤CT显示病变骨质呈显著膨胀性改变,骨破坏边缘及内部见多发骨嵴及粗大骨小梁,高密度骨针呈放射状、簇状或不规则排列。结论骨内脂肪瘤及血管脂肪瘤CT及MRI表现均以脂肪成分为主要特征,骨旁脂肪瘤CT以骨化性团块为特征,骨血管脂肪瘤骨质膨胀性改变较骨内脂肪瘤显著,病灶边缘及中心以多发骨嵴及粗大骨小梁为特征。