绿脓杆菌制剂对人肝癌MHCC97L细胞增殖和侵袭能力的影响

Objective To investigate the effects of Pseudomonas aeruginosa vaccine (PA) on proliferation and invasiveness of the hepatocellular carcinoma cell line MHCC97L with metastatic potential. Methods Proliferation, growth curve, plate efficiency, flow cytometry, transwell invasion assay, cell motility assay, scarification test, vascular endothelial growth factor (VEGF) and matrix metalloproteinase-2 (MMP2) protein activity were evaluated after cells were treated with PA at various concentrations. Results PA can inhibit the proliferation and plate efficiency of MHCC97L cell markedly in a dose-dependent manner. The IC50 of cells treated with PA for 48 h and 72 h was 3.1 ×108/ml and 1.9 × 108/ml, respectively. The doubling time increased and plate efficiency decreased gradually when cells treated with 0.5 × 108/ml, 1 × 108/ml and 2 × 108/ml PA (P＜0.01). PA could induce cell cycle arrest at the G1 phase in a dose-dependent manner by flow cytometric analysis. The average amount of invading cell per field in cell invasion assay and motility assay were 4. 8 ± 1.3 and 8. 8±2.2 when cells treated with 1× 108/ml PA, which was significantly lower than that of control group (8. 6±2. 1 and 15. 6±1.2 ) (P＜0.01) Scarification test showed that the metastatic ability of cells treated with 1 × 108/ml PA significantly lower than that in the control group. Comparison between cells treated with 1 × 108/ml PA and control group, no remarkable difference was found regarding expression of VEGF and MMP2 in supernatant of cell culture. Conclusion PA can inhibit proliferation and plate efficiency of HCC cell line MHCC97L, which is in part mediated by the cell cycle arrest at the G1 phase. PA could inhibit invasiveness of HCC cell line MHCC97L, which is unrelated to the VEGF and MMP2 protein activity.     

腹腔镜下膀胱尿道单针连续吻合法动物模型的建立和应用

Objective To develop and evaluate a porcine model for training the single needle running suture method of laparoscopie urethrovesical anastomosis(LUA). Methods Twenty minipigs with mean weight of 30kg were general anaesthetized with Sumianxin solution 0. 1 ml/kg intramuscularly. Pneumoperitoneum was created by insufflation of carbon dioxide by a veress needle inserted through the umbilicus. One 10mm port and two 5mm ports were positioned after the establishment of pneumoperitoneum. The intestine was used as "bladder". The procedures were completed with the single needle running suture method of laparoscopic urethrovesical anastomosis. Six trainees performed the LUA procedure based on the models during a laparoscopic training course, following the technique used in the operation room. The learning curve was analyzed by operative time. Results The porcine model for laparoscopic training was established successfully and 3 LUAs could be performed on each pig. Each trainee performed 10 LUAs based on the models during the training course of laparoscopic urology. The operative time declined from (55.3±10. 4)min initially to (22.4±4.8)min (P＜0. 01) after the training course. At the end of training, all trainees could accomplish a watertight LUR procedure on the model. Conclusions The establishment of the training model is feasible. The trainees could acquire the skills necessary to perform LUA in vivo based on this model. The model provides a platform for training the basic techniques of LUA procedures.     

螺旋CT三维成像技术在后腹腔镜下保留肾单位手术前的应用价值

漏尿发生. 结论 螺旋CT三维成像技术能清晰重建肾脏解剖构像,为选择后腹腔镜下NSS治疗肾肿瘤提供可靠依据.     

多原发大肠癌的临床诊断和外科治疗

目的 探讨多原发大肠癌的临床特点、诊断和治疗方法.方法 对1998-2007年收治的47例同时性多原发大肠癌和20例异时性多原发大肠癌患者的临床病理资料进行回顾性分析.结果 67例多原发大肠癌中同时性多原发大肠癌47例(占70%),其中直肠癌占39%.Dukes分期(以病理分期最晚的一个为准):A期6例,B期22例,C期15例,D期4例.67例中伴有淋巴结转移者20例,肠腔内伴有息肉者21例,无息肉者26例.行全结肠切除术3例,结肠次全切除术10例,根治性手术加肠段联合切除术34例.异时性多原发癌20例(占30%),共有癌灶44个,以结肠癌多见,共31例(占70%),二原发癌17例,三原发癌2例,四原发癌1例.与原发癌间隔时间7个月至19年,其中<2年者7例,2～5年者5例,>5年者8例.20例异时性多原发癌的初发癌均行根治性手术,14例二次癌和2例三次癌均行根治性切除术.同时性多原发大肠癌和异时性多原发癌初发癌根治术后5年生存率分别为74%和78%.结论 治疗大肠癌切忌满足于单一肠段或单个肿瘤的诊断和治疗,应利用结肠镜对全肠道进行仔细地检查,以便于及时发现大肠的多原发肿瘤;在根治性切除肠道肿瘤的同时应尽可能多地保留残存的正常肠道,从而更好地改善患者术后的生活质量.     

移植肾功能延迟恢复患者尿脱落细胞中穿孔素和粒酶B mRNA定量检测的意义

目的 探讨尿脱落细胞中穿孔素mRNA、粒酶B mRNA检测在移植肾功能延迟恢复(DGF)诊断中的应用价值. 方法 同种异体肾移植DGF患者24例.男15例,女9例.平均年龄37岁.移植肾活检病理分析病因为输尿管梗阻2例、静脉血栓1例、急性环孢素(CsA)中毒3例、急性肾小管坏死(ATN)7例、ATN合并临界改变2例、ATN合并急性排斥反应(AR)3例、AR 6例.移植术后1～14 d留取24例患者73份晨尿标本,应用竞争RT-PCR方法 对患者尿脱落细胞中穿孔素和粒酶B mRNA进行定量检测,应用SPSS13.0软件进行数据分析,mRNA水平用自然对数值表示.结果 24例患者尿脱落细胞中穿孔素和粒酶B mRNA水平在急性CsA中毒、ATN和其他组分别为(-0.76±0.35)、(-0.89±0.30)、(-0.90±0.15)fg/μg和(-0.53±0.22)、(-0.41±0.17)、(-0.73±0.23)fg/μg,组间厕两比较差异均无统计学意义(P>0.05);ATN合并AR组与单纯AR组分别为(1.20±0.39)、(11.13±0.33)fg/μg和(1.07±0.30)、(1.01±0.19)fg/μg,2组比较差异无统计学意义(P>0.05);ATN合并AR、单纯AR组与急性CsA中毒、ATN和其他组比较差异有统计学意义(P<0.001);2例ATN合并临界改变患者尿中穿孔素和粒酶B mRNA平均值分别为1.22、0.97 fg/μg,与ATN合并AR、单纯AR组相近. 结论 DGF患者尿脱落细胞中穿孔素和粒酶BmRNA水平检测有助于DGF的病因诊断,可作为临界改变是否需要进一步干预性治疗的指标之一.     

醛固酮合酶和11-β羟化酶基因多态性与原发性醛固酮增多症发病风险的相关性研究

目的 研究醛固酮合酶基因(CYP11B2)和11-β羟化酶基因(CYP11B1)多态性与原发性醛同酮增多症(PA)发病风险之间的相关性.方法 对醛固酮瘤(APA)和特发性醛崮酮增多症(IHA)患者(APA 134例,IHA 45例)及正常人群(118名)中CYP11B2和CYP1181基因的5个多态性位点进行检测.其中,intron 2采用2个独立的PCR反应,其余位点均采用Taqman探针法.采用Haploview 4.0、SNPassoc 1.5-3和Haplo.stats 1.3.8软件分析CYPllB2和CYP1181基因多态性与PA的关系.结果 所选位点均获成功检测,APA和IHA组中rs6410位点的A等位基因频数显著高于对照组(P=1.09×10-5,P=0.015).与对照组相比,APA组中rs6410位点AA基因型和AG基因型比例增高(P=2.19×10-4),而IHA组中rs6410位点AA基因型和AG基因型仅在年龄、性别和体质量指数校对后比例增高(OR=4.06,95%CJ 1.31～12.66;OR=2.41,95%CI 1.02～5.72).APA组中发现1个易感单体型AAAWT(OR=1.44,95%CI 1.19～1.76),IHA组中发现3个易感单体型AAAWT、AGGWT和AGAWC(OR=1.55,95%CI 1.23～1.96;OR=1.49,95%CI 1.17～1.89;OR:1.40,95%CI 1.04～1.88).同时在APA组中发现1个保护性单体型GGAWT(OR:0.73,95%CI 0.55～0.97).结论 CYP11B2和CYP11B1基因多态性与APA和IHA的发病显著相关.     

解剖性后腹腔镜下肾上腺切除术的阶段性培训方法

目的 建立解剖性后腹腔镜下肾上腺切除术(ARA)的培训方法,评估其安全性和有效性. 方法 5名无开放性肾上腺手术经验的青年医师接受3个阶段腹腔镜培训:①模拟箱训练定向转移、切割及缝合,共300 h;②动物模型训练辨认和游离组织、切割止血及精细缝合打结,共120 h;③临床实践:初期仅为导师扶持腹腔镜,再独立完成简单的腹腔镜手术如肾囊肿去顶术,最后在导师指导下独立完成30例ARA.学员作ARA病例的选择由导师负责,与导师最初的选择一致(除外肾上腺嗜铬细胞瘤).记录患者术前资料包括性别、年龄、体质量指数、肿瘤位置、肿瘤大小、肿瘤病理类型,记录5名学员共完成150例ARA(学员组)总的中转开放率、手术时间、估计出血量、平均住院时间、手术并发症,并与导师最初完成30例ARA(导师组)的相应参数进行比较.采用SPSS 12.0软件进行数据处理,计数、计量资料分别采用x2和t检验(除手术时间为偏态分布,采用非参数检验).结果学员完成ARA患者的术前资料均与导师组患者资料相匹配(P值均>0.05).每名学员都成功实施了30例ARA,无中转开放手术者.学员组150例平均手术时间为82(59～133)min,短于导师组30例的手术时间132(73～230)min(P<0.01).学员组150例ARA的估计出血量为(62.2±22.0)ml,平均住院时间(4.8±1.3)d,与导师组30例ARA的估计出血量(63.9±21.1)ml和平均住院时间(4.5±1.4)d比较,差异无统计学意义(P>0.05).学员组与导师组围手术期并发症发生率比较差异无统计学意义(8.0%和13.3%,P>0.05),但学员组术中轻微并发症发生率(1.3%)低于导师组(10.0%,P<0.05). 结论 阶段性培训方法能安全、有效地使青年泌尿外科医师掌握ARA.     

南京地区人群CYP2E1基因多态性及吸烟、饮酒与前列腺癌易感性

目的:探讨Ⅰ相代谢酶细胞色素P450 2E1(CYP2E1)基因RsaⅠ和PstⅠ位点多态性及吸烟、饮酒习惯与前列腺癌(PCa)发病风险的关系,并探讨基因与生活习惯在PCa发病中的联合作用。方法:采用PCR-RFLP技术检测109例原发性PCa患者及202例年龄匹配的男性非肿瘤患者外周血CYP2E1基因RsaⅠ和PstⅠ多态位点的基因型。结果:深吸烟(OR=2.29,95%CI:1.28～4.09)、重度吸烟史(OR=1.81,95%CI:1.02～3.22)等生活习惯为PCa易感因素。CYP2E1 C1/C1基因型与PCa易感性有显著相关(OR=1.71,95%CI:1.04～2.82),且与饮酒的联合作用明显与PCa易感性相关(OR=2.21,95%CI:1.06～4.59)。重度吸烟人群中,携带CYP2E1易感基因型(C1/C1)与不吸烟且携带非易感基因型(C1/C2或C2/C2)个体相比PCa的发病风险显著增高(OR=2.80,95%CI:1.20～6.56)。结论:携带CYP2E1易感基因型(C1/C1)并有烟酒嗜好者PCa的发病风险显著增高,且与烟草的暴露呈显著剂量反应关系。     

腹腔镜肾盂成形术的多模培训模式

目的：介绍一项腹腔镜肾盂成形术（Laparoscopic Pyeloplasty,LP）的多模培训模式,并评价其安全性、可行性和有效性。方法：该多模培训模式包括模拟器、动物模型和手术室培训三部分,5位具有不同开放肾盂成形术和腹腔镜经验的学员参加了这项培训,我们对5个学员参加培训过程中的相关数据和学员独立完成手术的5组LP患者的围手术期数据进行了评价。结果：5位学员均成功地完成了整个培训,并成功地独立完成了5例LP。所有患者手术均未转开放手术,均未输血,无患者死亡。模拟器培训和动物模型培圳所需的时间,学员1（3周,7天）比学员5（4周,8天）少,学员4（2周,6天）比学员2（3周,8天）和学员5（4周,8天）少。在手术室培训中,各学员训练每个手术步骤所需的手术例数相当。5位学员独立完成手术的5组患者手术时间、估计失血量、术后住院时间和围手术期并发症的差异均无统计学意义（P〉0．05）。结论：该项多模培训模式是安全、可行和有效的,并适合于没有开放肾盂成形术和腹腔镜经验的学员进行LP培训。     

后腹腔镜下根治性肾切除术及其应用解剖学研究

目的 总结后腹腔镜下根治性肾切除术,并对镜下肾周区域相关解剖结构进行研究.方法 2006年1月至2008年3月行后腹腔镜下根治性肾切除术85例.肿瘤位于左肾38例,右肾47例;肿瘤直径2.5～10.5 cm,平均(5.5±1.7)cm;T1N0Mo 74例,T2:N0M0 11例.根据肾筋膜外切除原则,在"两个间隙、上下两极间"分离切除患肾,腹侧为肾旁前间隙(后腹膜与肾前筋膜之间),背侧为腰肌前间隙(肾后筋膜与腰肌筋膜之间),上极达膈下,下方到髂窝.术中辨认重要的解剖结构:后腹膜及其返折、肾前筋膜、侧锥筋膜、肾后筋膜、腰大肌及重要血管. 结果 除1例因与周围粘连严重中转开放手术外,其余均获得成功.手术时间50～165 min,中位数65 min.估计失血量25～600ml,中位数58 ml.术中腹膜破裂5例,肾蒂周围小血管损伤出血6例,无重要脏器及大血管损伤.术后平均随访10(2～25)个月,均无瘤生存,无局部复发或发生穿刺通道种植转移. 结论 后腹腔镜下根治性肾切除术时,熟悉重要的镜下解剖结构有利于提高手术安全性、减少副损伤.