含精氨酸-甘氨酸-天冬氨酸肽的白细胞介素-24突变体真核表达载体的构建及其体外抗肝癌作用

目的 构建含精氨酸-甘氨酸-天冬氨酸(RGD)肽的白细胞介素(IL)-24突变体(RGD-IL-24)真核表达质粒,观察其对肝癌细胞的抑制作用.方法 利用重叠PCR技术在IL-24cDNA中插入GGT三个碱基,获得RGD-IL-24 cDNA.将其克隆入真核表达载体pcDNA3.1(+),构建pcDNA3.1/RGD-IL-24.将pcDNA3-1/RGD-IL-24及作为对照的pcDNA3.1/IL-24转染肝癌细胞HepG2,逆转录-聚合酶链反应(RT-PCR)检测IL-24 mRNA表达;Western blot检测IL-24蛋白表达;噻唑蓝(MTT)检测细胞增殖;FITC-Annexin V染色检测细胞凋亡;Western blot检测bax、bcl-2表达.结果 测序证明pcDNA3.1/RGD-IL-24构建成功.RT-PCR、Western blot证实转染pcDNA3.1/RGD-IL-24的肝癌HepG2细胞表达IL-24.pcDNA3.1/RGD-IL-24、pcDNA3.1/IL-24处理组HepG2细胞存活率分别为(0.382±0.064、0.563±0.038),凋亡细胞阳性率分别为(0.346±0.049、0.163±0.087),两组差异均有统计学意义.Western blot证实pcDNA3.1/RGD-IL-24促进bax表达、抑制bcl-2表达作用强于pcDNA3.1/IL-24.结论 RGD-IL-24真核表达质粒不仅不影响IL-24表达,且能显著增强IL-24诱导肝癌细胞凋亡及抑制肿瘤生长作用.     

肿瘤Ki-67抗原HLA-A2限制性细胞毒性T淋巴细胞表位的鉴定

Objective To identify the HLA-A2-restricted CTL epitopes encoded by antigen segment of Ki-67 for preparation of tumor vaccines. Methods Two epitope prediction databases ( BIMAS and SYFPEITHI) were used to predict the epitopes of Ki-67, and 7 peptides were synthesized as the candi-dates. The prediction was evaluated by peptide-binding test. ELISPOT was used to inspect the IFN-γ secre-ted by CTLs in order to reflect the immunogenicity of the peptides. Results The purities of synthetic pep-tides and positive control peptide were all above 95%. Peptide-binding test displayed that LQGETQLLV (280-288) of Ki-67 could bind with HLA-A2 molecule more strongly. ELISPOT assay displayed that LQGETQLLV could induce activation of specific CTLs. Conclusion LQGETQLLV (280-288) might be the HLA-A2-restricted CTL epitope of Ki-67 antigen.     

表达端粒酶逆转录酶siRNA的溶瘤腺病毒对裸鼠肾癌移植瘤的治疗作用

Objective To investigate the antitumor effect of oncolytic adenovirus armed with small interference RNA targeting hTERT gene for renal cancer therapy. Methods Nude mice were divid-ed randomly into 4 groups (8 mice/group),and were treated by intratumoral injections of ZD55-hTERT ( an oncolytic adenovirus armed with small interference RNA targeting hTERT gene) ,ZD55-EGFP ( an on-colytic adenovirus) and Ad-hTERT (replication-defective adenovirus armed with small interference RNA targeting hTERT gene) with three consecutive daily at 7 × 108 pfu/day or treated with PBS as a control. The expression of E1A and hTERT, and apoptosis of tumor xenografts were assessed by immunohistochemi-cal technique at the 7th day after injections. The tumor volume was measured at the 50th day after injec-tions. Results The tumor volume in ZD55-hTERT treatment group ( 124.1±27.5) was significantly less than that in ZD-EGFP (499.8±77.1 ) and Ad-hTERT ( 609.0±102.5 ) treatment groups. The E 1A pos-itive expression in ZD55-hTERT treatment group was significantly higher than that in Ad-hTERT treatment group. The hTERT positive expression in ZD55-hTERT treatment group was significantly lower than that in Ad-hTERT treatment group. ZD55-hTERT treatment of tumor xenografts resulted in an increased apoptotie cell death as compared with ZD55-EGFP and Ad-hTERT treatment. Conclusion The antitumor effect of ZD55-hTERT was more potent than oneolytie adenovirus ZD55-EGFP and Ad-hTERT.     

Solitaire AB支架在辅助栓塞颅内宽颈动脉瘤中的临床应用

目的评价Solitaire AB支架在辅助栓塞颅内宽颈动脉瘤中的临床应用价值。方法应用Solitaire AB支架治疗颅内宽颈动脉瘤25例,其中后交通动脉瘤15枚,眼动脉段动脉瘤4枚,颈内动脉海绵窦段动脉瘤1枚,大脑中动脉瘤1枚,椎动脉动脉瘤4枚。结果 25例中,完全栓塞22例,瘤颈残留1例,部分栓塞2例。术中动脉瘤破裂1例,术后脑积水行脑脊液分流术1例,弹簧圈移位1例。随访未有再出血病例。造影随访12例,复发1例。结论应用Solitaire AB支架在辅助栓塞颅内宽颈动脉瘤具有较高的临床价值,值得临床推广。     

过氧化物还原酶Ⅱ在胃癌中的表达及临床病理意义

背景与目的:过氧化物还原酶Ⅱ(peroxiredoxinⅡ,PrxⅡ)是具有过氧化物酶活性的蛋白,相关研究提示其参与多种恶性肿瘤的发生、发展.该实验通过研究人类胃癌组织及细胞中的PrxⅡ的表达情况,分析其与胃癌临床病理特征的关系,探讨其与胃癌发生、发展及预后的关系.方法:采用实时荧光定量聚合酶链反应(real-time fluorescent quantitative polymerase chain reaction,RTFQ-PCR)和蛋白[质]印迹法(Western blot)检测45例胃癌患者的癌组织及相应的癌旁组织、胃正常细胞(GES-1)及胃腺癌细胞(MGC-803、MKN-45和MKN-28)的PrxⅡmRNA及蛋白表达情况.应用免疫组织化学S-P法检测胃癌组织芯片中116例胃癌患者癌组织及癌旁组织的PrxⅡ的表达量,分析PrxⅡ表达与胃癌临床病理特征的关系并做生存分析.结果:RTFQ-PCR和Western blot检测结果显示,胃癌组织中PrxⅡmRNA及蛋白表达水平明显高于相应的癌旁组织(P<0.05).胃癌细胞中PrxⅡmRNA及蛋白表达水平明显高于胃正常细胞(P<0.01).免疫组织化学结果显示,胃癌组织中PrxⅡ蛋白的表达阳性率(76.7％)同样明显高于相应癌旁组织(30.1％,P<0.01).PrxⅡ蛋白表达与胃癌的肿瘤大小、分化程度、浸润深度、TNM分期及淋巴结转移密切相关(P<0.05),而与患者的性别、年龄、肿瘤部位及远处转移无关(P>0.05).PrxⅡ蛋白高表达者的生存时间要显著低于低表达者(P<0.01),PrxⅡ是影响胃癌预后的独立危险因素.结论:PrxⅡ促进胃癌的发生、发展,是胃癌的不良预后因素,其研究可能为胃癌的治疗提供新的靶点.     

血管腔内治疗婴幼儿布加综合征 1 例报告

目的探讨婴幼儿布-加综合征(BCS)的血管腔内治疗。方法回顾分析1例婴儿期诊断为BCS并行血管腔内治疗患儿的临床资料。结果患儿,女,23个月,反复腹胀3月余,经血管超声、磁共振成像检查确诊为BCS。明确诊断后行血管腔内治疗,术后闭塞血管内血流通畅,尿量增加,腹胀明显减轻。结论婴幼儿BCS罕见,病情复杂,行血管腔内治疗有效。     

经不同途径置入覆膜支架封堵气管瘘的临床分析

目的探讨经气管或食管置入覆膜支架封堵气管瘘的疗效。方法13例气管瘘患者,男9例,女4例,年龄55～83岁,平均60．3岁,其中食管癌性狭窄伴气管瘘8例,食管癌术后胃吻合口伴气管瘘3例,食管癌术后胸腔胃气管隆突瘘1例,肺癌术后左主支气管残端胸膜瘘1例。根据原发病变的性质、狭窄范围、瘘口部位的不同,在数字减影血管造影（DSA）监视下经食管或气管途径置入个体化设计覆膜支架封堵瘘口。结果10例经食管置入覆膜支架10枚,其中管状覆膜支架8枚,下端大喇叭口覆膜支架2枚;3例经气管置入覆膜支架4枚,其中管状覆膜支架2枚,倒“Y”型覆膜支架1枚,“子弹头”残端瘘支架1枚。所有患者呛咳、呼吸困难和憋喘等症状迅速缓解,碘水造影显示瘘口封堵完全,CT检查显示支架贴壁良好。结论经气管或食管置入个体化覆膜支架治疗气管瘘,能迅速封堵瘘口和解除临床症状,简单、安全,值得推广。     

联合支架置入术治疗恶性气管、食管和上腔静脉复合狭窄

目的探讨联合支架置入术在恶性气管、食管、上腔静脉复合狭窄急诊治疗中的临床疗效。方法4例恶性肿瘤造成气管、食管、上腔静脉狭窄患者,男3例,女1例,年龄50～77岁,平均67岁。经螺旋CT气管、上腔静脉三维重建和食道造影检查,明确气管、食管、上腔静脉狭窄的部位和程度。根据患者原发疾病的特点和病情症状的缓急,在数字减影血管造影透视下先后置入气管和食管支架2例,先后置入气管和上腔静脉支架2例。结果所有支架均一次性置入成功,共置入气管支架4枚（其中气管倒Y型一体化支架1枚）,食管支架2枚,上腔静脉支架3枚。患者置入气管和食管支架后,呼吸和吞咽困难症状明显改善;置入气管和上腔静脉支架后,呼吸困难和头面部、上肢肿胀症状明显改善。脉氧饱和度由高流量吸氧时的70％～80％提高至自然呼吸时的90％～95％。结论联合支架置入术治疗恶性气管、食管和上腔静脉复合狭窄操作简单、安全,能明显提高患者的生活质量,是行之有效的急救措施之一。     

经颈静脉介入治疗不同类型急性髂股静脉血栓

目的 评价经颈静脉介入治疗不同类型急性髂股静脉血栓形成(IF-VT)的疗效.方法 38例急性IF-VT患者来自徐州医学院附属医院,全部经颈静脉插管行尿激酶溶栓治疗.结果 左侧IF-VT或伴血栓蔓延至下腔静脉16例;左侧IF-VT伴左髂总静脉闭塞17例;左侧IF-VT合并左髂总静脉狭窄或闭塞伴血栓蔓延至下腔静脉3例;双侧IF-VT伴左髂总静脉狭窄或闭塞2例.血栓完全溶解33例,部分溶解5例.20例随访2～18个月,血栓复发2例.结论 经颈静脉介入治疗不同类型急性IF-VT是一种安全、有效的治疗方法.     

布加综合征合并肝静脉血栓介入治疗的疗效观察

目的 评价布加综合征合并肝静脉血栓介入治疗的临床效果.方法 25例布加综合征合并肝静脉血栓形成的患者,均施行经导管尿激酶溶栓术、球囊扩张术和(或)支架置入术,术后随访采用肝脏超声检查,观察肝静脉、下腔静脉通畅情况及血栓有无复发.介入治疗前后肝静脉-右心房、下腔静脉-右心房压差的比较采用配对t检验.结果 治疗成功23例,其中血栓完全溶解18例,部分溶解5例,肝静脉及下腔静脉血流通畅,肝静脉-右心房压差由术前平均(29±7)cm H2O(1 cm H2O＝0.098 kPa)下降至术后平均(8±3)cm H2O(t=13.7,P＜0.01),下腔静脉-右心房压差由术前平均(19±4)cm H2O下降至术后平均(5±2)cm H2O(t=13.3,P＜0.01);不成功2例.23例患者随访1～42个月,平均(18±10)个月,死亡1例,肝静脉再狭窄2例,经再次球囊扩张治疗成功,其余20例无肝静脉再狭窄及血栓复发.结论 布加综合征合并肝静脉血栓的介入治疗可取得较好临床效果.

Abstract：

Objective To evaluate the effect of interventional therapy for Budd-Chiari syndrome with hepatic vein thrombosis. Methods Twenty-five patients with Budd-Chiari syndrome complicated with hepatic vein thrombosis underwent catheter-directed urokinase thrombolysis, balloon dilation and/or stent placement. During follow-up, re-thrombosis and patency of the recanalized hepatic vein and inferior vena cava were evaluated by liver ultrasound. The pressure gradient of hepatic vein-right atrium or inferior vena cava-right atrium before and after interventional treatment was compared with paired t-test. ResultsTechnical success was obtained in 23 patients. Complete resolution and partial resolution of the thrombi were accomplished in 18 cases and 5 cases, respectively. The recanalized hepatic veins and inferior vena cava were patent. The mean pressure gradient of hepatic vein-right atrium dropped from (29±7) cm H2O to (8±3) cm H2O (1 cm H2O＝0.098 kPa) after the interventional treatment (t=13.7,P<0.01). The mean pressure gradient of inferior vena cava-right atrium dropped from (19±4) cm H2O to (5±2) cm H2O after the interventional treatment (t=13.3, P<0.01). Failures occurred in 2 patients. Over the follow-up period of 1 to 42 months[(18±10) months]after interventional treatment in the 23 patients, one late death occurred. Restenoses of hepatic veins were found in 2 patients, which were all redilated successfully. Neither restenosis of hepatic vein nor recurrence of thrombosis was found in the other 20 patients. Conclusion Interventional therapy could be effectively performed for the treatment of Budd-Chiari syndrome with hepatic vein thrombosis.