microRNA-133a在复发性肝细胞癌中差异表达及其生物学功能研究

目的 探讨miR-133a在不同复发时间间隔复发性肝癌中的表达差异,并研究其对肝癌细胞侵袭、迁移、增殖等生物学功能的影响。方法 通过miRNA芯片筛选复发性肝细胞癌病例差异表达的miRNA;将miR-133a mimic及inhibitor转染入人肝癌细胞株SMMC-7721,通过CCK8检测细胞增殖能力,Transwell实验检测细胞迁移及侵袭能力;利用生物信息学方法预测miR-133a的可能靶基因。结果 芯片结果发现miR-133a在早期复发肝癌样本中表达明显升高;转染miR-133a的mimic或inhibitor入肝癌细胞SMMC-7721后,细胞增殖能力无显著差异(P>0.05);转染mimic后,细胞的侵袭及迁移能力明显增强(P<0.05),而转染inhibitor后,细胞的侵袭及迁移能力明显减弱(P<0.05)。结论 miR-133a的表达在不同复发间隔的肝细胞癌组间有显著差异,在短期复发肿瘤组织内明显升高;miR-133a可以促进肿瘤细胞的侵袭和迁移,而对增殖无明显作用,提示miR-133a水平升高可能通过促进肿瘤细胞侵袭从而增加肝细胞癌肝内转移复发风险。     

巧用一次性输液器瓶塞穿刺针保护套

近来，我们巧用一次性输液器瓶塞穿刺针保护套，以解决袋装液体与瓶装液体对一次性输液器排气孔的要求不同所带来的问题，取得满意效果。现报告如下。     

IL-10和IL-12在结直肠癌肝转移中作用的临床研究

目的探讨IL-10、IL-12在结直肠癌肝转移中作用。方法分别收集自2008年1月到2010年6月健康成年人、结直肠癌术前、术后和术后肝转移患者外周血,采用ELISA法测定外周血清IL-10、IL-12水平,采用FACS Calibur3.0流式细胞仪测定T淋巴细胞及其亚群比例,进行各组结果统计学分析。结果结直肠癌术后肝转移患者外周血清IL-10水平显著高于健康成年人、结直肠癌术后患者(P<0.05),结直肠癌术后患者外周血清IL-10水平显著低于术前患者(P<0.05);结直肠癌术前、术后和术后肝转移患者外周血清IL-12水平显著低于健康成年人,结直肠癌术前、术后患者之间以及术后和术后肝转移患者之间外周血清IL-12水平存在统计学差异(P<0.05);结直肠癌术后肝转移患者外周血T淋巴细胞总数及Th细胞比例数显著低于健康成年人(P<0.05),Ts细胞比例数显著高于健康成年人(P<0.05)。NK细胞在结直肠癌术后肝转移患者显著低于健康成年人(P<0.05)。结论不同临床时期结直肠癌患者存在细胞免疫或体液免疫严重障碍,纠正机体细胞因子和免疫状态紊乱,不仅能够改善机体免疫功能,可能有助于控制结直肠癌的发生、发展。     

雌激素受体β在乳腺癌中的表达

目的:探讨雌激素受体(ERβ)在乳腺癌表达情况以及其与乳腺癌的关系.方法:选取经根治性手术治疗的乳腺癌患者52例为研究对象.采用免疫组织化学法测定乳腺癌组织中ERβ的表达.根据ERβ表达情况将患者分成ERβ阳性组和阴性组,并结合临床病理指标和生物学指示作对比分析.结果:ERβ表达与Ki67表达负相关(P=0.019),ERβ蛋白的表达与月经状况,肿瘤大小, TNM分期,组织学类型无关,与其他生物学指标P53、C-erbB-2和GST-π表达无关.结论:ERβ表达状况对乳腺癌患者预后的判断有一定价值,为独立的预后影响因素.     

阻抑素在胃肠癌中的表达情况及其临床意义

目的检测阻抑素（PHB）在胃肠癌中的表达情况,探讨PHB对胃肠肿瘤诊断的价值。方法采用免疫组织化学法检测PHB在胃肠癌癌组织和癌旁组织中的表达情况。结果PHB在胃肠癌组织的表达量明显高于其相应癌旁组织的表达量,差异有极显著性（P〈0．01）;PHB在低分化胃癌、肠癌组织中的表达阜明显高于中分化癌组织,差异有极显著性（P〈0．01）。结论PHB在胃肠癌中特异性高表达,检测胃癌、肠癌组织中PHB的表达对胃癌、肠痛及其分化程度具有潜在诊断价值,为胃肠癌的临床诊断和治疗奠定了实验基础。     

马钱子碱免疫纳米微粒的研制及药代动力学特点

背景：由于马钱子碱具有剧毒、难溶于水、静脉应用治疗窗窄、中毒量与治疗量接近等缺点,限制了其临床应用于肝癌等恶性肿瘤的治疗.目的：研制马钱子碱免疫纳米微粒,观察马钱子碱免疫纳米微粒体内药物代谢特点.方法：利用阴离子聚合和化学改性技术制备羧基化聚乙二醇-聚乳酸嵌段共聚物,采用超声乳化技术制备羧基化聚乙二醇-聚乳酸嵌段共聚物马钱子碱纳米微粒,化学偶联技术将马钱子碱纳米微粒与抗人甲胎蛋白单克隆抗体结合,研制具有免疫靶向特点的药物制剂马钱子碱免疫纳米微粒.结果与结论：马钱子碱免疫纳米微粒外观圆整,大小较均一,平均粒径（249±77） nm,Zeta 电位（-18.7±4.19） mV.马钱子碱包封率（76.0±2.3）%,载药量（5.6±0.2）%.马钱子碱免疫纳米微粒在释放介质中24 h 累积释放80%以上,48 h 释放完全.马钱子碱免疫纳米微粒在体内代谢过程属于非房室模型,半衰期为（15.69±3.77） h,显著长于马钱子碱半衰期（P 〈 0.01）.提示实验成功研制了免疫靶向药物马钱子碱免疫纳米微粒,马钱子碱免疫纳米微粒在体内代谢过程属于非房室模型,表现出明显的缓释性.     

马钱子对人肝癌细胞生长的影响及其作用机制

目的 探讨马钱子对人肝癌细胞生长影响及作用机制.方法 体外培养人肝癌细胞SMMC-7721,分别加入5％、10％、20％的马钱子药物血清,噻唑蓝(MTT)比色法测定对人肝癌细胞SMMC-7721的生长抑制作用.20％马钱子药物血清作用肝癌细胞SMMC-77210、24、48 h后,分别收集细胞,提取蛋白和总RNA,应用Westem blot和荧光定量聚合酶链反应(PCR)检测Fas、Cyclin D1、PCNA蛋白和mRNA表达.结果 20％马钱子药物血清对人肝癌细胞72 h的生长抑制率为(54.94±8.19)％,与5％、10％浓度抑制率(19.928±7.653)％、(29.020 ±6.100)％比较差异有统计学意义(F=18.23,P＜0.01).药物血清作用24、48 h后,肝癌细胞SMMC-7721 Fas蛋白表达分别为(0.302±0.009)、(0.399±0.021),与对照组(0.226±0.022)比较,差异有统计学意义(F=68.25,P＜0.05);药物血清作用48 h后,肝癌细胞SMMC-7721 PCNA蛋白表达为(0.7457±0.0386),与对照组、24 h(0.8529±0.0099、0.9016±0.0139)比较,差异有统计学意义(P＜0.05);药物血清作用24、48 h后,肝癌细胞SMMC-7721 Cyclin D1 mRNA表达分别为0.045 680 ±0.038 130、0.026 714±0.019934,与对照组(0.145246±0.048543)比较,差异有统计学意义(F=8.671,P＜0.05);药物血清作用24、48 h后,肝癌细胞SMMC-7721 Cyclin D1蛋白表达,PCNA、Fas mRNA表达变化差异无统计学意义(F =68.25,P＞0.05).结论 马钱子具有抑制人肝癌细胞增殖作用,其抑制效应与药物血清浓度成正相关;通过上调肝癌细胞Fas蛋白和下调PCNA蛋白及Cyclin D1 mRNA表达,是马钱子抑制肝癌细胞增殖的机制之一.     

钽棒植入治疗早期股骨头坏死的护理体会

对25例早期股骨头坏死(Fical分期:I期10例,Ⅱ期15例)患者行钽棒植入术。术前充分准备,进行心理护理、健康教育;术后密切观察生命体征,加强体位护理及系统的康复锻炼,做好并发症的观察及预防。结果手术均获成功,无1例发生并发症。随访期内除1例左髋术后出现持续性疼痛外,其余24例术后髋部疼痛、活动受限均有明显改善。X线片复查股骨头骨坏死分期未加重。提示钽棒植入治疗早期股骨头坏死近期安全有效,全方位的护理是保证治疗效果的关键。     

水通道蛋白1在急性坏死性胰腺炎大鼠胰腺组织的表达及其意义

Objective To study the expression of aquaporin 1 ( AQP1 ) in pancreas and its pathogenic role in rats with experimental acute necrotizing pancreatitis (ANP). Methods Forty-eight male Sprague-Dawley rats were randomly divided into two groups : control group ( n = 24 ) and ANP group ( n = 24). Six rats in each group were sacrificed at 3,6,12 and 18 h after induction of experimental models. Quantity of ascites and levels of serum amylases were measured at each time point. Serum AQPI was determined by ELISA. Pathological changes of pancreatic tissues were examined. Capillary permeability in pancreatic tissues was tested by Evans Blue (EB) extravasation method. AQPI expression in pancreatic tissues was detected by immunohistochemieal staining, Western blot and fluorescence quantitative polymerase chain reaction (FQ-PCR). Results (1) Serum amylase level at 3,6,12, and 18 h in control group was (1308±759) ,(1077±508), (1325±761),(1328±762) U/L,and that in ANP group was (9102± 2199), (8799±1634), (9398±1473), (9484±862) U/L respectively. The concentration of EB in pancreatic tissues at each time point in control group was (205.61±32.99), (141.46±27.18 ), (96.94± 26.79), (61.43±24.82) mg/L, and that in ANP group was ( 273.59±23.47 ), ( 253.51±31.68 ), (221.15±73.68 ), (185.28±42.35) ,respectively. There was significandy difference between two groups. Serum AQP1 level in control group at each point was ( 74.08±11.80), (78.49±9.06 ), (75.77± 7.37 ), ( 72.75±13.87 ) mg/L, and that in AN P group was (73.29±9.61 ), ( 62.85±7.28 ), (62.07± 4.39 ), (46.33±11.91 ) mg/L, respectively ( P < 0.01 ). ( 2 ) Protein expression of AQPI in pancreatic tissues detected by immunohistochemical staining in control group at each time point was 114.13±7.92, 122.39±7.99,145.98±6.48,113.98±6.48, and that in ANP group was 80.07±14.89,110.54± 4.45,103.77±10.48,99.18±6.95;Protein expression of AQP1 in pancreatic tissues detected by Western blot in control group at each time point was 1.19±0.33,1.02±0.25,0.90±0.33,1.06±0.20,and that in ANP group was 0.83±0.11,0.96±0.21,0. 58±0.28,0.72±0.14, respectively ( P < 0.05 ). ( 3 ) The mRNA level of AQP1 gene expressed in pancreas in control group at each time point was 0.91±0.22,1.01±0.83,0.48±0.23,0.61±0.51,and that in ANP group was 2.13±0.63,2.02±1.40, 2.07±0.86,2.49±2.47,respectively (P<0.01).Condusion The expression of AQP1 wasdown-reg-ulated in pancreas in rats with ANP,which might could play an important role in the pathogenesis of capil-lary leak syndrome.     

水通道蛋白1在急性坏死性胰腺炎大鼠胰腺组织的表达及其意义

Objective To study the expression of aquaporin 1 ( AQP1 ) in pancreas and its pathogenic role in rats with experimental acute necrotizing pancreatitis (ANP). Methods Forty-eight male Sprague-Dawley rats were randomly divided into two groups : control group ( n = 24 ) and ANP group ( n = 24). Six rats in each group were sacrificed at 3,6,12 and 18 h after induction of experimental models. Quantity of ascites and levels of serum amylases were measured at each time point. Serum AQPI was determined by ELISA. Pathological changes of pancreatic tissues were examined. Capillary permeability in pancreatic tissues was tested by Evans Blue (EB) extravasation method. AQPI expression in pancreatic tissues was detected by immunohistochemieal staining, Western blot and fluorescence quantitative polymerase chain reaction (FQ-PCR). Results (1) Serum amylase level at 3,6,12, and 18 h in control group was (1308±759) ,(1077±508), (1325±761),(1328±762) U/L,and that in ANP group was (9102± 2199), (8799±1634), (9398±1473), (9484±862) U/L respectively. The concentration of EB in pancreatic tissues at each time point in control group was (205.61±32.99), (141.46±27.18 ), (96.94± 26.79), (61.43±24.82) mg/L, and that in ANP group was ( 273.59±23.47 ), ( 253.51±31.68 ), (221.15±73.68 ), (185.28±42.35) ,respectively. There was significandy difference between two groups. Serum AQP1 level in control group at each point was ( 74.08±11.80), (78.49±9.06 ), (75.77± 7.37 ), ( 72.75±13.87 ) mg/L, and that in AN P group was (73.29±9.61 ), ( 62.85±7.28 ), (62.07± 4.39 ), (46.33±11.91 ) mg/L, respectively ( P < 0.01 ). ( 2 ) Protein expression of AQPI in pancreatic tissues detected by immunohistochemical staining in control group at each time point was 114.13±7.92, 122.39±7.99,145.98±6.48,113.98±6.48, and that in ANP group was 80.07±14.89,110.54± 4.45,103.77±10.48,99.18±6.95;Protein expression of AQP1 in pancreatic tissues detected by Western blot in control group at each time point was 1.19±0.33,1.02±0.25,0.90±0.33,1.06±0.20,and that in ANP group was 0.83±0.11,0.96±0.21,0. 58±0.28,0.72±0.14, respectively ( P < 0.05 ). ( 3 ) The mRNA level of AQP1 gene expressed in pancreas in control group at each time point was 0.91±0.22,1.01±0.83,0.48±0.23,0.61±0.51,and that in ANP group was 2.13±0.63,2.02±1.40, 2.07±0.86,2.49±2.47,respectively (P<0.01).Condusion The expression of AQP1 wasdown-reg-ulated in pancreas in rats with ANP,which might could play an important role in the pathogenesis of capil-lary leak syndrome.