结直肠癌中Galectin-9的表达及其意义

目的：探讨Galectin-9基因和蛋白在结直肠癌中的表达及其临床意义。方法：运用荧光实时定量PCR及免疫组织化学染色方法检测45例手术切除的结直肠癌标本和20例正常结直肠上皮组织中Galectin-9基因和蛋白的表达，分析Galectin-9的表达与结直肠癌临床病理特征的关系。结果：Galectin-9基因在结直肠癌组织中表达低于正常结直肠上皮组织（P〈0．05），Galectin-9蛋白在结直肠癌组织中表达阳性率为62％，在正常结直肠上皮组织中表达阳性率为90％，差异具有统计学意义（P〈0．05）。Galectin-9的低表达与结直肠癌的远处转移相关（P〈0．05），与患者的年龄、性别、肿瘤浸润深度、淋巴结转移、肿瘤分化程度、肿瘤分期及肿瘤部位无显著相关。结论：Galectin-9在结直肠癌中低表达，并且与肿瘤的远处转移相关，有可能成为结直肠癌判断预后的标志物以及基因治疗的潜在靶点。     

老年患者腹腔镜结直肠手术的安全性研究

Objective To compare the outcomes of laparoscopic colorectal surgery in elderly (≥ 70 years) and younger (< 70 years) patients, and to evaluate the safety of laparoscopic colorectal surgery in elderly. Methods From Aug. 2007 to Dec. 2007, 25 patients with colorectal cancer undergone laparoscopic colorectal surgery were studied prospectively. Ten patients aged 70 years or older (Elder Group), and 15 patients less than 70 years (Younger Group). The parameters of perioperative hemodynamics, blood chemistry, and intraabdominal organs perfusion were recorded periodically during operation and compared between the two groups. Results Elder group had a higher ASA degree (P<0.01). Mean age, height, weight, the classification of the disease, operative time, blood loss and complication were not significantly different between the two groups. The HCO3 value and PaCO2 value increased significantly intraoperatively, while the pH value decreased significantly. These parameters returned to basal line at the end of operation, with the decreased electrolytes, especially the kaliopenia and hypocalcemia. The CVP increased significantly at the beginning of operation and after fluid expansion, and decreased postoperatively. Gastric mucosal pH(pHi) decreased when the operation began. The partial pressure of gastric mucesa (PgCO2) increased significantly at the end of operation.Other parameters of hemodynamics including HR, CI, and SI were not significantly different during operation. The CVP of elder group was higher than that of younger group at 5 min after the beginning of the operation, while the MAP of elder group was lower than that of younger at 15 min after the end of the operation. Other parameters of hemodynamics including oxygenation, pedusion, and blood chemistry were not significantly different. Conclusions For laparoscopic colorectal surgery, no significant adverse response attributes to elderly patients. The compensation capacity of the elderly is lower. With the reasonable anesthesia administration, the adverse response can be controlled effectively.     

Galectin-9基因不同亚型表达载体的构建

目的构建galectin-9基因不同亚型的真核表达载体。方法应用RT-PCR方法从人正常大肠上皮组织中扩增出galec-tin-9三种剪接体的全长片段,分别将片段连接到质粒载体pIRES2-EGFP上,经测序证实插入片段正确后,载体转染结肠癌LoVo细胞,再应用RT-PCR和Western blot方法证实转染效果。结果测序结果证实载体上插入的galectin-9三种剪接体碱基序列正确,同时载体成功转染LoVo细胞,并成功表达三种剪接体蛋白。结论成功构建了galectin-9基因不同亚型的真核表达载体,为研究galectin-9的多种功能创造了条件。     

腹腔镜胃局部切除术的临床应用

目的 评价腹腔镜胃局部切除术的临床应用价值.方法 回顾性分析23例腹腔镜胃局部切除手术患者的临床资料,包括肿瘤大小和位置、手术时间、术中失血量、术后排气时间、术后住院天数、手术并发症、术后病理结果及随访情况.结果 23例患者的胃局部切除术均在腹腔镜下成功完成,无中转开腹手术.其中8例实施腹腔镜锲型胃局部切除术,15例实施腹腔镜经胃腔黏膜下病灶切除术.手术切除肿瘤直径(2.8±1.3)cm,手术时间(82.2±35.5)min,术中失血量(26.5±15.3)mL,手术切口长度(3.1±1.1)cm,术后恢复排气时间(2.1±0.9)d,术后住院天数(7.8±2.0)d.2例(8.7%)在接受腹腔镜经胃腔黏膜下病灶切除术后出现胃腔内渗血,经非手术治疗后痊愈.中位随访时间12个月(2～45个月),未发现肿瘤复发和戳口种植.结论 腹腔镜胃局部切除术是治疗良性胃肿瘤安全、可行及微创的手术方法.     

两种不同腔镜腹股沟疝修补术CO2充气对血流动力学与呼吸功能的影响

目的:本临床研究通过比较经腹腔腹膜前腹股沟疝修补(TAPP)术和完全腹膜外腹股沟疝修补(TEP)术在不同的操作空间充入CO2气体时对病人呼吸和循环功能的影响,以论证腔镜腹股沟疝修补术的安全性.方法:将2005年1月至2005年6月,本中心将收治的腹股沟疝病人随机分成两组:20例采用经腹腔腹膜前腹股沟疝修补手术(TAPP组),20例采用完全腹膜外腹股沟疝修补手术(TEP组).按不同的操作空间充入CO2气体,观察并比较两组手术相关指标及病理生理指标.结果:TAPP组和TEP组病人存性别、年龄、体重指数、疝类型、手术时间、住院天数、术后疼痛分数等方面.差异无统计学意义(P＞0.05);TAPP组和TEP组病人术后均出现心率减慢和血压上升,呼气末CO:分压值上升,pH值下降,PaCO2和HCO3值上升,术后5 min的指标与术前比较差异有统计学意义(P＜0.05).随着手术时间的延长,各指标的变化进一步加大.拔管后各指标迅速趋于恢复术前水平,但与术前指标比较,差异仍有统计学意义(P＜0.05):TAPP组和TEP组的组间比较,各指标之间的差异在各个时间段均无统计学意义(P＞0.05).结论:TAPP和TEP手术均对病人的呼吸、循环产生一定影响,出现CO2蓄积、酸中毒、血压上升及心率减慢等表现.通过合理调节,可以将这些指标控制在安全的范围内,两种手术都是安全的.     

Galectin-9基因不同亚型对结肠癌LoVo细胞株体外黏附侵袭能力的影响

目的 构建galeetin-9基因不同亚型的真核表达载体,观察galectin-9对结肠癌LoVo细胞体外黏附侵袭能力的影响.方法 构建galectin-9基因不同亚型的真核表达载体后,用脂质体法转染结肠癌LoVo细胞,逆转录-聚合酶链反应(RT-PCR)和Western blot方法验证转染效果.LoVo细胞分别在转染galectin-9、转染galectin-9+抗体(5 mg/L)和转染galectin-9+β-L糖(30 mmol/L)条件下进行体外细胞-基质黏附实验和体外细胞侵袭实验.结果 测序结果证实载体上插入的galectin-9三种剪接体碱基序列正确,同时载体成功转染LoVo细胞.体外细胞-基质黏附实验中,转染空载体,转染galectin-9L,galectin-9M和galectin-9S组平均吸光度分别为:0.74±0.04、1.14±0.12、1.12±0.10和1.15±0.13,表明galectin-9促进LoVo细胞黏附于细胞外基质(P＜0.05).galectin-9抗体和β-乳糖可抑制该作用,但转染galectin-9L和galectin-9S LoVo细胞的黏附作用受β-乳糖抑制更明显(P＜0.05).体外侵袭实验未发现galectin-9对LoVo细胞侵袭力有影响.结论 galectin-9促进LoVo细胞与细胞外基质的黏附,这可能与结肠癌侵袭转移相关,但其三种亚型发挥作用的机制不全相同.     

老年人ERCP的安全性和有效性探讨

目的通过比较不同年龄段患者中内镜逆行胰胆管造影（endoscopic retrograde cholangiopancreatography，ERCP）的应用情况，分析和探讨ERCP在老年患者中应用的有效性和安全性。方法2006年1月～2007年12月，本中心进行ERCP操作的患者301例，按年龄分组，其中≥70岁的患者（老年组）102例，〈70岁的患者（年轻组）199例。记录患者ASA分级、基础疾病、ERCP操作以及并发症情况，对结果进行统计分析。结果301例患者进行了313次ERCP操作。老年组患者ASA分级比年轻组高，差异有统计学意义。老年组患者临床症状中黄疸和胆道感染占30．4％．年轻组患者非特异症状占85．9％，差异有统计学意义。术前生化指标在两组间差异无统计学意义。ERCP操作成功率为96．7％（96．1％vs97．0％），取石成功率94．6％（94．8％vs94．4％），结石取尽率94．9％（94．5％vs95．1％）．41例患者采用了预切开术（12．7％vs14．1％），差异无统计学意义。ERCP结果显示，胆总管结石是最常见的诊断（56．9％vs54.3％），恶性肿瘤（20．6％）和十二指肠乳头周围憩室（37．3％）多见于老年患者。并发症14例，老年组5例（4．9％），年轻组9例（4．5％），两组间差异无统计学意义。结论老年人进行ERCP操作能够取得和年轻人一样的疗效．尽管老年人ASA分级较高，但ERCP并发症并不高于年轻人。ERCP在老年患者中的应用是安全有效的。     

腹腔镜低位直肠保肛术中直肠脱出技术应用的初步研究

目的：探讨直肠脱出技术在腹腔镜低位保肛术中的应用价值。方法：回顾分析2005年10月至2009年1月运用直肠脱出技术行腹腔镜低位直肠前切除术的20例低位直肠肿瘤病人的临床资料,探讨其手术操作步骤及术后恢复情况、肿瘤根治性效果和随访结果。结果：所有病例均未发生术中严重并发症和手术死广,无中转开腹手术：平均手术时间为（122＋801min,术中平均出m（50±70）mL,病人术后平均排气时间、留置导尿管时间及术后住院天数分别为（2．2±1．1）d、（4．9±2．4）d和（11．0±4．2）d。,每例病人的平均清扫淋巴结总数为（15±4）枚,肿瘤距下切缘平均距离为（2．2±1．1）cm。所有病人均获随访,中位随访时间19（2-36）个月,未发现局部复发和远处转移,排便功能恢复均较满意。结论：直肠脱出技术运用于腹腔镜低位直肠保肛术安全有效,可作为低位直肠肿瘤保肛术的有效技术。     

Demethylation of the γ-synuclein gene CpG island in colorectal cancer and its clinical significance

Objective To explore the relationship between γ-synuclein gene expression and CpG island demethylation in colorectal cancer (CRC), and the relationship between the demethylation and clinicopathological factors of CRC. Methods The expression of γ-synuclein mRNA was examined in 30 pairs of tumor tissues and tumor-matched non-neoplastic adjacent tissues (NNAT) by RT-PCR.CRC cell lines including COLO205, LoVo, and SW480 were used and treated with a demethylating agent, 5-aza-2'-deoxycytidine (5-aza-C). Before and after the treatment, the expression of γ-synuclein mRNA in the cells was determined by RT-PCR, and bisulfite sequencing PCR was also used to analyze methylation status of CpG island. The methylation status of γ-synuclein was then examined in 67 CRC samples and 30 NNAT samples by nested methylation-specific PCR (NMSP) and real time methylationspecific PCR (real-time MSP). The relationship between the demethylation of γ-synuclein in CRC and clinicopathological factors was analyzed. Results The mean γ-synuclein mRNA expression was 0.66±0.34 in CRC samples, which was much higher than 0.45±0.26 in NNAT samples (P=0.011). 5-aza-C could induce expression and demethylation of γ-synuclein in COLO205, LoVo and SW480 cells. Γ-Synuclein gene was demethylated in 80.0%(24/30) of the CRC samples and 50.0%(15/30) of the NNAT samples.The demethylated status of γ-synuclein was much higher in CRC samples than that in NNAT samples (P=0.030), and was significantly correlated with clinical stage, lymph node involvement, and distant metastasis of CRC (P＜0.05). Conclusion The upregulation of γ-synuclein expression in CRC is primarily attributed to the demethylation of CpG island, which may be used as a marker for prognosis.