蛋白激酶D1及其甲基化在胃癌中的表达研究

目的 探讨蛋白激酶D1（PKD1）在胃癌组织和胃癌细胞中的表达,及其甲基化在胃癌发病中的作用.方法 采用RT-PCR方法检测胃癌组织中PKD1 mRNA的表达;体外培养胃癌细胞系BGC-823和MGC-803,采用Western blot检测PKD1甲基化特异性阻断剂5-杂氮-2＇-脱氧胞苷（5-Aza-CdR）处理前后PKD1蛋白的表达.结果 与癌旁组织相比,PKD1在胃癌组织中的表达明显下降（0.318±0.036比0.908±0.041,P＜0.01）.5-Aza-CdR处理后,BGC-823和MGC-803细胞中PKD1蛋白表达均显著增高（均P＜0.05）.结论 PKD1甲基化可导致胃癌细胞PKD1表达下降,可能参与了胃癌的发生和进展.     

胃泌素及其受体拮抗剂对 BGC-823细胞骨架微丝结构的作用

目的 探讨胃泌素及其受体拮抗剂对人胃癌BGC-823细胞系细胞骨架微丝结构的作用。方法 在体外用胃泌素和受体拮抗剂处理胃癌BGC-823细胞系,制备光镜和电镜标本并观察细胞骨架,结果 胃泌素组细胞核增大,胞浆内微丝量明显减少,微丝部位不定,不呈束结构紊乱;而胃泌素和受体拮抗剂组胞浆内边缘微丝、中间微丝、核周微丝均明显增多,排列规则并呈束,其结构和数量均大体相似于对照组,结论 胃泌素能增强BGC-823细胞系的运动,有促进肿瘤细胞转移的作用,而受体拮抗剂可抑制胃泌素的这种作用。     

P27kipl基因过度表达诱导人胃癌细胞系凋亡机制的研究

目的研究P27kipl基因转染对人胃癌细胞系BGC-823的诱导凋亡作用及其可能机制。方法构建含人P27kipl基因的逆转录病毒真核表达载体,将外源性P27kipl基因转染人胃癌细胞系BGC-823,应用Westernblot、流式细胞分析术(FCM)等方法,检测P27kipl蛋白在胃癌细胞内的表达、诱导凋亡作用及其对bcl-2基因表达的影响。结果外源性P27kipl蛋白在胃癌细胞中获稳定表达,胃癌细胞凋亡,bcl-2蛋白表达水平降低。结论P27kipl的过度表达可通过对bcl-2表达的调控显著促进胃癌细胞凋亡,提示P27kipl基因转染可成为胃癌的有效治疗方法。     

肿瘤坏死因子相关凋亡诱导配体通过促进死亡相关蛋白3表达抑制人胃癌细胞株的生长

目的：观察肿瘤坏死因子相关凋亡诱导配体（TRAIL）对人胃癌细胞株的生长抑制作用及其作用前、后人胃癌细胞株中死亡相关蛋白3（DAP3）表达情况，探讨DAP3在TRAIL抑制人胃癌细胞株生长中的作用。方法：选择不同浓度（0、50、100、200ng／m1）TRAIL作用于人胃癌细胞株BGC-823和HGC-2748h后，用酸性磷酸酶法测定各组细胞株生长抑制情况，确定TRAIL的有效浓度：取有效浓度的TRAIL处理人胃癌BGC-823和HGC-27细胞株48h后，分别用Westem印迹法和RT—PCR检测TRAIL对DAP3蛋白和DAP3mRNA表达的影响。结果：①酸性磷酸酶法测得100ng／mlTRAIL可有效抑制人胃癌细胞株BGC-823和HGC-27生长。②Western印迹法未能在人胃癌细胞株BGC-823和HGC-27中测得DAP3表达．而100ng／mlTRAIL处理后48h，在BGC-823和HGC-27中均可测得DAP3表达：同样．RT—PCR法测得100ng／mlTRAIL处理48h后．两株胃癌细胞株中DAP3mRNA表达显著升高。结论：TRAIL能有效抑制人胃癌细胞株BGC-823和HGC-27的生长．其机制与促进DAP3表达有关。     

P27^kip1基因过度表达诱导人胃癌细胞系凋亡机制的研究

目的：研究P27^kipl基因转染对人胃癌细胞系BGC-823的诱导凋亡作用及其可能机制。方法：构建含人P27^kipl基因的逆转录病毒真核表达载体，将外源性P27^kipl基因转染人胃癌细胞系BGC-823，应用Western blot、流式细胞分析术（FCM）等方法，检测P27^kipl蛋白在胃癌细胞内的表达、诱导凋亡作用及其对bcl-2基因表达的影响。结果：外源性P27^kipl蛋白在胃癌细胞中获稳定表达，胃癌细胞凋亡，bcl-2蛋白表达水平降低。结论：P27^kipl的过度表达可通过对bcl-2表达的调控显促进胃癌细胞凋亡，提示P27^kipl基因转染可成为胃癌的有效治疗方法。     

RNA干扰抑制yrdC蛋白表达对人胃癌细胞株BGC-823体外增殖能力的影响

目的:构建针对人yrdC基因siRNA的重组腺病毒Ad-shyrdC并观察其转染人胃癌细胞株BGC-823后对细胞体外增殖能力的影响。方法:筛选高效抑制yrdC蛋白表达的RNAi位点,并构建表达siRNA的重组腺病毒,转染人胃癌细胞株BGC-823后以Western印迹法鉴定细胞yrdC蛋白表达的变化;同时通过MTT法、流式细胞术分别测定BGC-823细胞增殖活性和增殖指数的变化。结果:成功构建了抑制人yrdC基因siRNA的重组腺病毒Ad-shyrdC;转染BGC-823细胞后可明显抑制yrdC蛋白表达,并使所测定的细胞MTT值和细胞增殖指数提示BGC-823细胞增殖活性下降(P<0.05)。结论:腺病毒介导的RNAi能有效地抑制BGC-823细胞中yrdC蛋白表达,并降低其增殖活性。     

A14125I-胰岛素与H22肝癌细胞胰岛素受体结合的研究

目的体外分析H22肝癌细胞与正常肝细胞膜的胰岛素受体表达,探讨胰岛素作为受体介导靶向治疗载体的可行性及临床应用前景.方法用氯氨T法制备A14125I-胰岛素,电泳分离纯化标记物并用自身置换比放射性测定和过量受体结合实验法鉴定标记物质量,再制备H22肝癌细胞及正常小鼠肝细胞,进行体外受体结合实验,分别测定125I-胰岛素与小鼠H22肝癌及正常肝细胞膜胰岛素受体结合的Kd值及Bmax值以及其竞争结合曲线.125I-胰岛素受体结合的Kd值及Bmax值用t检验分析,饱和结合曲线用Scatchard分析.结果小鼠H22肝癌细胞及正常小鼠肝细胞胰岛素受体Bmax值分别为(5.6±1.1) pmol/106细胞和(3.2±0.8) pmol/106细胞,高亲和力Kd值为(1.8±0.6) nmol/L及(2.1±0.9) nmol/L,低亲和力Kd值为(32.0±10.7) nmol/L及(37.0±12.3) nmol/L.癌细胞胰岛素受体Bmax值明显高于正常肝细胞(P＜0.05).受体结合实验表明,125I-胰岛素与H22肝癌及正常细胞的结合具有特异性.结论 125I-胰岛素与H22肝癌及正常小鼠肝细胞膜受体的结合具有高度特异性,H22肝癌细胞较正常小鼠肝细胞表达胰岛素受体增加,提示胰岛素可作为抗癌载体通过与肝癌细胞表面受体结合而介导肝癌的靶向治疗.     

人肝癌细胞膜胰岛素受体的研究及临床意义

目的　研究临床上用胰岛素作为载体通过受体介导对肝癌进行靶向治疗的可行性及其临床价值。方法　以A14 12 5I 标记胰岛素 ,取人肝癌细胞及癌旁组织 ,制备细胞膜 ,测定细胞膜胰岛素受体密度并评价其受体结合活性。结果　A14 12 5I 胰岛素比放射性为 (46 1± 1 2 )TBq/mmol,放射化学纯度 >98% ;人肝癌及癌旁组织细胞膜胰岛素受体Bmax分别为 (1 9± 0 6 )pmol/mg蛋白及(1 4± 0 6 )pmol/mg蛋白 ,高亲和力Kd(解离常数值 )值为 (2 1± 0 6 )nmol及 (2 2± 0 8)nmol,低亲和力Kd值为 (2 5± 8)nmol及 (2 7± 8)nmol。人肝癌细胞膜胰岛素受体表达高于癌旁组织 (最大结合值Bmaxt=0 0 32 ,P <0 0 5 )。结论　A14 12 5I 胰岛素与人肝癌细胞膜胰岛素受体具有特异性的结合力     

一种特异性靶向性淋巴细胞对荷瘤裸鼠的治疗观察

目的观察癌胚抗原(CEA)阳性靶向性淋巴细胞对胃癌细胞体内及体外靶向性治疗作用。方法从健康人外周血中分离单个核细胞(PBMC),通过脂质体lipofectamine 2000介导的细胞转染将重组真核表达载体anti-CEA-sc Fv-CD3ζ-pc DNA3.0介导的细胞转染导入PBMC,以获取CEA阳性靶向性淋巴细胞(以下简称靶淋巴细胞);将不含anti-CEA-sc Fv-CD3ζ-pc DNA3.0融合基因片段的pc DNA3.0真核表达载体转染入PBMC,获取的淋巴细胞简称为空载体淋巴细胞。将上述两种淋巴细胞分别与CEA阳性KATOⅢ胃癌细胞和CEA阴性BGC-823胃癌细胞共培养,观察不同靶淋巴细胞与胃癌细胞共培养(24 h或36 h)后的识别情况或其对胃癌细胞凋亡的影响。并将上述已获取的两种淋巴细胞分别通过尾静脉注入荷CEA阳性KATOⅢ胃癌细胞裸鼠和荷CEA阴性BGC-823胃癌细胞裸鼠体内,以观察其对荷胃癌裸鼠肿瘤生长情况的影响。结果 (1)靶淋巴细胞与胃癌细胞共培养24 h时,靶淋巴细胞对KATOⅢ胃癌细胞的识别率为72.3%,淋巴细胞对KATOⅢ胃癌细胞的识别能力较强;其对BGC-823胃癌细胞识别率为7.8%,其对BGC-823胃癌细胞识别能力较弱。(2)当靶淋巴细胞、空载体淋巴细胞分别与KATOⅢ和BGC-823胃癌细胞分别共培养36 h时,靶淋巴细胞+KATOⅢ胃癌细胞组的凋亡率明显高于空载体淋巴细胞+KATOⅢ胃癌细胞组(P=0.032),而靶淋巴细胞+BGC-823胃癌细胞组和空载体淋巴细胞+BGC-823胃癌细胞组的凋亡率比较差异无统计学意义(P=0.118)。(3)在观察40 d内,靶淋巴细胞+KATOⅢ胃癌细胞组的肿瘤体积明显小于空载体淋巴细胞+KATOⅢ胃癌细胞组(F=5.010,P0.01)和空白对照组(F=7.560,P0.01);靶淋巴细胞+BGC-823胃癌细胞组与空载体淋巴细胞+BGC-823胃癌细胞组间比较差异无统计学意义(F=1.210,P0.05)。靶淋巴细胞+KATOⅢ胃癌细胞组的肿瘤体积明显小于靶淋巴细胞+BGC-823胃癌细胞组(F=4.982,P0.01)。结论 CEA阳性靶向性淋巴细胞可特异性促进CEA阳性胃癌细胞凋亡,抑制荷CEA阳性胃癌细胞裸鼠肿瘤的生长。     

CCL22-CCR4轴在胃癌腹膜乳斑转移的机制研究

目的 探讨巨噬细胞衍生因子MDC/CCL22-CCR4反应轴在胃癌腹膜乳斑转移中的作用.方法 用RT-PCR和Western blot方法检测CCR4在5种胃癌细胞系中的表达;MTT法测定不同浓度CCL22对胃癌细胞株BGC-823增殖率的变化;Transwell小室体外侵袭实验检测CCL22对胃癌细胞株BGC-823趋化活性的影响;免疫组化检测标本中CCR4表达;单变量分析CCR4表达与胃癌临床病理参数的关系.结果 CCR4mRNA和CCR4蛋白在5种胃癌细胞系中均有表达.CCL22可以促进BGC-823细胞的增殖和趋化.CCR4在胃癌中有表达,CCR4表达水平与胃癌分化程度相关(P<0.05),而与患者的年龄、肿瘤大小、位置、分期以及淋巴结转移等均无关(P>0.05).结论 CCL22-CCR4反应轴可能在胃癌腹膜乳斑转移中起作用.CCR4可能成为预防和治疗胃癌腹膜转移的潜在靶点.     

国产与进口6%羟乙基淀粉130/0.4注射液容量治疗疗效和安全性的对比研究

目的 与进口6%羟乙基淀粉130/0.4注射液进行对比性研究,评价国产6%羟乙基淀粉130/0.4注射液的有效性及安全性.方法 患者随机分为研究组123例,对照组117例,术中输注研究药物或对照药物10 ml·kg-1·h-1,输注时间 2 h,期间观察患者生命体征,记录液体出入量与合并用药,比较术前和输注后血常规、血电解质、凝血功能、肝肾功能、尿常规指标变化;并记录术中不良事件发生与处理.结果 两组患者用药期间BP、HR、SpO2均能维持正常,BP和HR的变化率两组间差异无统计学意义.两组患者术中出入量、应用血管活性药物及应用血液制品人数与用量组间差异无统计学意义.输注后两组Hct、Hb下降幅度一致,血K+、Na+略低于应用前,血Cl-高于应用前,血糖有升高,PT、APTT时间延长,TT时间略缩短.肝功能ALT、AST超过正常值1.5倍的两组各有4例.肾功能BUN、Cr研究组有11例,对照组7例高于正常.结论 国产6%羟乙基淀粉130/0.4注射液用于全麻下非心脏外科手术患者术中容量治疗扩容效果安全可靠,与进口6%羟乙基淀粉130/0.4注射液作用相似.     

The Central Role of the Nose in the Face and the Psyche: Review of the Nose and the Psyche

Background According to statistics released by the American Society of Plastic Surgeons in 2006, rhinoplasty is one of the most sought after aesthetic surgeries by ethnic patients and teenagers. It also is the most requested aesthetic operation by patients with body dysmorphic disorder. The psychosocial aspect of rhinoplasty is undeniable. Tagliacozzi in 1567 and Joseph more than a century ago were already aware of this aspect. Methods Using the terms “rhinoplasty,” “patients selection,” “psychological aspect,” and psychological outcome,” 30 studies were selected through searches of the MEDLINE, PUBMED, and EMBASE databases,. This review aimed to analyze how the most acknowledged experts of psychology, facial plastic surgery, and plastic surgery who have worked on the psychological outcome for rhinoplasty during the past century considered the nose–psyche relationship and the influence of rhinoplasty at the psychological level. Results The link between rhinoplasty, psychology, and social environment has been discussed by many important authors during the past century. All of them, independently of their field of study, have stressed that it is critical for surgeons to be aware of their responsibility regarding both the physical and emotional levels. There is evidence that an official preoperative interview is lacking. Conclusions To recognize the importance of rhinoplasty’s psychological implications, it is critical to make a good selection of patients seeking this operation and to have a good outcome. To this end, the preoperative interview is fundamental. Surgeons should be competent at both the psychological and surgical levels.     

腹腔镜联合内镜微创手术治疗胆囊结石合并胆总管结石的临床观察

目的探讨腹腔镜联合内镜微创手术治疗胆囊结石合并胆总管结石的疗效。方法回顾性分析95例胆囊结石合并胆总管结石病人行腹腔镜联合内镜微创手术治疗的临床资料。结果本组63例先行十二指肠镜下乳头括约肌切开术(endoscopic sphincterotomy,EST)治疗,其中59例成功行EST+腹腔镜胆囊切除术(laparoscopic cholecystectomy,LC),但术中并发十二指肠乳头少量出血2例,术后发生胆道感染1例,出现可疑十二指肠漏1例;4例EST取石失败后1周内改开腹手术,术后并发腹腔感染1例,胆漏1例。23例顺利行LC+腹腔镜胆总管探查取石术,其中腹腔镜胆囊管探查取石5例,腹腔镜胆总管切开取石18例。9例因疑诊胆总管结石而先行LC,术后2~4 d再行EST。术后随访6~12个月,均未出现反流性胆管炎、乳头狭窄等并发症,无胆管结石残留。结论腹腔镜联合内镜微创手术治疗胆囊结石合并胆总管结石效果满意。     

Reconciling the capability approach and the ICF

The literature on applications of the so-called “Capability Approach” of Amartya Sen and Martha Nussbaum is extensive, but it is only recently that some have argued that its application to the analysis of disability would be a great advantage over existing analyses, and in particular preferable to the model of functioning and disability found in the World Health Organization's International Classification of Functioning, Disability and Health (ICF). I argue here that care must be taken in this head-to-head comparison between the Capability Approach and ICF since the former is essentially a political-theoretical account of equalitarian justice, whereas the latter is a model of a classification system for describing disability that is explicitly neutral between any theory of distributive justice. Nonetheless, this paper argues that a careful comparison of the two approaches to the conceptualisation of disability reveals salient aspects of convergence that, arguably, point to a potential synergy between the Capability Approach as applied to disability and the ICF.     

Teamwork and the legal and ethical responsibility of the anaesthetist

PURPOSE OF REVIEW: Anaesthetists are members of the operating team. Although the surgeons usually consider themselves to be the leaders of the team, all members, including the anaesthetist, have their own legal, ethical and professional responsibilities. RECENT FINDINGS: Good communication and sharing responsibility are characteristics of teamwork. Many factors can affect team functioning: training differences, social status, differences in responsibility, gender, age and culture, among others. An increasing number of individuals are involved in modern treatment due to limited working hours and increasing subspecialization, which complicates collaboration. Not all participants have the same perception of the team structure and its operation. Multidisciplinary, well designed training programmes are needed. SUMMARY: Anaesthetists have to be more aware of their position in the care team and pay more attention to their legal, ethical and professional responsibilities.     

尿动力学异常与慢性前列腺炎诊治关系的探讨

目的 :探讨下尿路功能性梗阻与慢性前列腺炎的关系。　方法 :通过尿动力学检查 ,证实 4 2例慢性前列腺炎病人存在下尿路功能性梗阻 ,主要表现有不稳定膀胱、括约肌痉挛、逼尿肌 外括约肌协同失调。 3 8例接受药物治疗 ,4例接受经尿道前列腺激光汽化术。　结果 :3 6例获得随访 ,治愈 2 5例 ( 69.4 % ) ,好转 10例 ( 2 7.8% ) ,无效 1例 ( 2 .8% )。　结论 :下尿路功能性梗阻在慢性前列腺炎的诊治中具有重要意义。