免钉合腹腔镜下350例完全腹膜外无张力疝修补术的应用

目的探讨腹腔镜下完全腹膜外疝修补术(totally extroperitoneal in-guinal hernia repair,TEP)应用的合理性。方法回顾性分析2002年11月至2009年7月行腹腔镜下完全腹膜外疝修补术共350例(356例次)患者的临床资料,总结术后并发症、复发率的发生情况。结果本组术后并发症发生率10.1%(36/356),复发率0.84%(3/356),平均手术时间60 min,术后平均住院时间3.5 d。结论TEP经由腹膜前间隙进入腹股沟区,采用补片覆盖耻骨肌孔,符合力学原理,不仅具有微创优势,而且比开放式无张力疝修补更具合理性。     

DNA甲基转移酶在肝癌中的表达及其临床意义

Objective To investigate the expression of DNA methyltransferases (DNMTs) in liver cancer and its clinical significance. Methods The specimens of liver cancer tissues, adjacent tissues, cirrhotic tissues and chronic hepatitis tissues were collected from 50 patients who received radical resection at the First Affiliated Hospital of Sun Yat-Sen University from July 2007 to April 2008. The mRNA and protein expressions of DNMT1,DNMT3a and DNMT3b in liver cancer tissues, adjacent tissues, cirrhotic tissues and chronic hepatitis tissues were detected by real-time quantitative PCR and immunohistochemical staining. The mRNA expression of DNMTs in the liver cancer tissues was compared with those in the adjacent tissues, cirrhotic tissues and chronic hepatitis tissues by using t test and Mann-Whitney U test. The correlation between the protein expression of DNMTs in the liver cancer tissue and the clinicopathological features was analyzed by chi-square test or Fisher exact test, and the tumor-free survival time was analyzed by using Kaplan-Meier method and the difference in tumor-free survival rate between different patients was analyzed by Log-rank test. Results The mRNA expressions of DNMT1, DNMT3a and DNMT3b in the liver cancer tissue were 2.57, 2.29 and 4.86 times higher than those in the adjacent tissues (t = 3.94, 2. 72, 4. 06, P ＜ 0.05 ). The mRNA expressions of DNMT1, DNMT3a and DNMT3b were 2.38,2.14 and 4.66 times higher than those in the cirrhotic tissues, and 6.12, 4.58 and 12.99 times higher than those in the chronic hepatitis tissues. The mRNA expressions of DNMT1, DNMT3a and DNMT3b in the liver cancer tissue were significantly higher than those in the cirrhotic tissues and chronic hepatitis tissues ( U = 587.5,730. 0,562.5; 65.5, 64.5, 71.0, P ＜ 0.05). The protein expression of DNMT1 was correlated with the size, number,TNM stages and vascular invasion of tumors ( x2 = 4.08, 5.95, 4.08, P ＜ 0.05 ). The protein expression of DNMT3a was correlated with the size, number and TNM stages of tumors (x2 = 4.08, 5.95, 4.08, P ＜ 0.05 ).The mean tumor recurrence time of patients with low expressions of DNMT1 and DNMT3a were 9.4 and 8.7 months, which were significantly longer than 5.0 and 3.2 months of those with high expressions of DNMT1 and DNMT3a (x2 =3.89, 9.91, P＜0.05). Conclusions DNMTs play an important role in hepatocarcinogenesis.High expressions of DNMT1 and DNMT3a are correlated with the postoperative recurrence of liver cancer, which are valuable prognostic factors for liver cancer.     

腹腔镜、胃镜双镜联合治疗胃部肿瘤的临床应用

目的评价腹腔镜、胃镜联合治疗胃部肿瘤的临床应用效果。方法行腹腔镜胃部肿瘤手术34例,术中结合胃镜行病变定位,其中行腹腔镜下全胃切除5例,腹腔镜下近端胃大部切除术3例,腹腔镜下远端胃大部切除术26例,腹腔镜下胃手术同时行腹腔镜胆囊切除术3例。结果 2例在腹腔镜下完成胃的分离、切除、吻合;32例先腹腔镜分离,然后通过上腹正中切一个5～8 mm切口,进行胃的切除、吻合,标本亦于此切口取出。无中转开腹病例。手术时间60-395分钟,术中出血30～200 mL,肠蠕动恢复时间28-60小时,下床活动时间36-48小时,进流质时间3-5天,本组病例无1例应用镇痛剂,未出现吻合口漏、腹腔出血、切口感染等并发症,仅有1例患者出现食管黏膜糜烂出血,治疗后痊愈。21例患者术后规律随访6月,未发现操作孔、小切口及腹腔内转移。结论腹腔镜、胃镜联合手术治疗胃部肿瘤具有微创、疼痛轻、胃肠功能恢复快、下床、住院时间短、切口小、对机体免疫功能影响小、术后恢复快等优点。     

胃肠间质瘤多学科综合治疗的价值及评价

胃肠间质瘤（gastrointestinal stromal tumor,GIST）是胃肠道最常见的间叶来源肿瘤。外科手术联合靶向药物综合治疗是GIST的合理治疗模式。建立多学科协作的诊疗制度,定期组织多学科病例讨论,形成稳定的治疗团队,能够快速提高疾病的诊疗水平,减少误诊,显著提高治疗效果。根据病人的具体情况制定科学、有效的个体化治疗方案。     

DNA甲基转移酶在肝门部胆管癌中的表达及临床意义

目的 探讨DNA甲基转移酶(DNMTs)在肝门部胆管癌中的表达及其临床意义.方法 收集1997年4月至2007年3月中山大学附属第一医院收治的111例肝门部胆管癌患者(胆管癌组)和39例胆总管囊肿患者(对照组)胆管组织标本,构建组织芯片,采用免疫组织化学方法检测DNMT1、DNMT3a、DNMT3b的蛋白表达水平;比较胆管癌组与对照组的胆管组织中DNMTs蛋白表达的差异;分析肝门部胆管癌组织中DNMTs蛋白表达与临床病理因素的关系,计数资料的比较采用x2检验或Fisher确切概率法,Kaplan-Meier法绘制生存曲线,生存情况分析采用Log-rank检验.结果 胆管癌组的DNMT1和DNMT3b高表达率分别为54.1％ (60/111)和47.7％ (53/111),显著高于对照组的28.2％ (11/39)和23.1％ (9/39),两组比较,差异有统计学意义(x2=7.740,7.240,P＜0.05).DNMT1的高表达与Bismuth-Corlette分型、T分期存在明显相关性(x2=12.200,17.800,P＜0.05);而DNMT3a在胆管癌组和对照组的高表达率比较,差异无统计学意义(x2=3.370,P＞0.05);DNMT3b的高表达则只与Bismuth-Corlette分型存在明显相关性(x2=8.300,P＜0.05).111例患者中66例行肝门部胆管癌切除术,其中42例获得随访.DNMT1低表达者的术后中位生存时间为23.9个月,明显长于DNMT1高表达者的11.8个月(x2=3.980,P＜0.05).结论 DNMT1和DNMT3b高表达在肝门部胆管癌发生、发展中起重要作用;DNMT1的表达水平与肝门部胆管癌根治性切除术后生存时间明显相关,可望成为有价值的预测因子.     

DNA甲基转移酶在肝癌中的表达及其临床意义

目的 探讨DNA甲基转移酶(DNMTs)在肝癌中的表达及其临床意义.方法 收集2007年7月至2008年4月中山大学附属第一医院行根治性切除的50例肝癌患者的肝癌组织、癌旁组织、肝硬化组织和慢性肝炎组织.采用RT-PCR和免疫组织化学方法检测肝癌组织及癌旁组织中DNMT1、DNMT3a、DNMT3b mRNA与蛋白表达水平,采用配对t检验和Mann-Whitney U检验比较肝癌组织与癌旁组织,肝硬化组织与慢性肝炎组织DNMTs mRNA表达的差异,x2检验和Fisher确切概率法分析肝癌组织DNMTs蛋白表达与临床病理因素的相关性,Kaplan-Meier法分析患者无瘤生存时间,不同患者无瘤生存时间的比较采用Log-rank检验.结果 肝癌组织中DNMT1、DNMT3a、DNMT3b mRNA的表达水平分别为癌旁组织的2.57、2.29和4.86倍(t=3.94,2.72,4.06,P＜0.05).肝癌组织中DNMT1、DNMT3a、DNMT3b mRNA的表达分别是肝硬化组织和慢性肝炎组织的2.38、2.14、4.66倍和6.12、4.58、12.99倍.DNMT1、DNMT3a、DNMT3b mRNA在肝癌组织的表达显著高于其在肝硬化组织和慢性肝炎组织中的表达(U=587.5、730.0、562.5,65.5、64.5、71.0,P＜0.05).DNMT1蛋白表达与肿瘤大小、数目、有无血管侵犯、TNM分期有关(x2=5.24,4.08,4.08,14.13,P＜0.05);DNMT3a蛋白表达与肿瘤大小、数目、TNM分期有关(x2=4.08,5.95,4.08,P＜0.05).DNMT1和DNMT3a低表达者平均肿瘤复发时间分别为9.4个月和8.7个月,显著长于高表达者的5.0个月和3.2个月,两者比较,差异有统计学意义(x2=3.89,9.91,P＜0.05).结论 DNMTs在肝癌发生、发展中起重要作用;DNMT1与DNMT3a高表达与肿瘤切除术后复发密切相关,可望成为肝癌预后的重要预测因子.

Abstract：

Objective To investigate the expression of DNA methyltransferases (DNMTs) in liver cancer and its clinical significance. Methods The specimens of liver cancer tissues, adjacent tissues, cirrhotic tissues and chronic hepatitis tissues were collected from 50 patients who received radical resection at the First Affiliated Hospital of Sun Yat-Sen University from July 2007 to April 2008. The mRNA and protein expressions of DNMT1,DNMT3a and DNMT3b in liver cancer tissues, adjacent tissues, cirrhotic tissues and chronic hepatitis tissues were detected by real-time quantitative PCR and immunohistochemical staining. The mRNA expression of DNMTs in the liver cancer tissues was compared with those in the adjacent tissues, cirrhotic tissues and chronic hepatitis tissues by using t test and Mann-Whitney U test. The correlation between the protein expression of DNMTs in the liver cancer tissue and the clinicopathological features was analyzed by chi-square test or Fisher exact test, and the tumor-free survival time was analyzed by using Kaplan-Meier method and the difference in tumor-free survival rate between different patients was analyzed by Log-rank test. Results The mRNA expressions of DNMT1, DNMT3a and DNMT3b in the liver cancer tissue were 2.57, 2.29 and 4.86 times higher than those in the adjacent tissues (t = 3.94, 2. 72, 4. 06, P ＜ 0.05 ). The mRNA expressions of DNMT1, DNMT3a and DNMT3b were 2.38,2.14 and 4.66 times higher than those in the cirrhotic tissues, and 6.12, 4.58 and 12.99 times higher than those in the chronic hepatitis tissues. The mRNA expressions of DNMT1, DNMT3a and DNMT3b in the liver cancer tissue were significantly higher than those in the cirrhotic tissues and chronic hepatitis tissues ( U = 587.5,730. 0,562.5; 65.5, 64.5, 71.0, P ＜ 0.05). The protein expression of DNMT1 was correlated with the size, number,TNM stages and vascular invasion of tumors ( x2 = 4.08, 5.95, 4.08, P ＜ 0.05 ). The protein expression of DNMT3a was correlated with the size, number and TNM stages of tumors (x2 = 4.08, 5.95, 4.08, P ＜ 0.05 ).The mean tumor recurrence time of patients with low expressions of DNMT1 and DNMT3a were 9.4 and 8.7 months, which were significantly longer than 5.0 and 3.2 months of those with high expressions of DNMT1 and DNMT3a (x2 =3.89, 9.91, P＜0.05). Conclusions DNMTs play an important role in hepatocarcinogenesis.High expressions of DNMT1 and DNMT3a are correlated with the postoperative recurrence of liver cancer, which are valuable prognostic factors for liver cancer.     

术前禁食时间对BALB/c裸鼠NK细胞杀伤活性的影响

目的 研究术前禁食时间长短对BALB/c裸鼠NK细胞杀伤活性的影响. 方法 按术前禁食时间分为24小时组、36小时组、48小时组,每组10只BALB/c裸鼠(5只为非禁食对照组),每只裸鼠行取脾术后处死,通过磁珠分选技术将裸鼠脾脏NK细胞分选出来,进而对NK细胞的杀伤活性进行检测.结果 禁食24小时和36小时后裸鼠的NK细胞活性均有不同程度地下降(分别为1.06％和2.2％),但差异无统计学意义(P＞0.05),在禁食48小时后可见裸鼠脾脏的NK细胞杀伤效应进一步下降(5.96％),差异有统计学意义(P=0.014). 结论 术前长时间禁食可降低裸鼠脾脏NK细胞的杀伤效应.     

微波消融灭瘤联合瘤内接种树突状细胞诱导特异性抗肝癌免疫

目的 探讨微波消融(MWA)灭瘤联合瘤内接种树突状细胞(DCs)诱导特异性抗肝癌免疫的效能.方法 采用GM-CSF联合IL-4体外培养C57BL/6小鼠骨髓来源的DCs,于第6天收集使用.建立C57BL/6小鼠皮下Hepa1-6肝癌模型,随机分为对照组、瘤内接种DCs组(DC组)、肿瘤微波消融组(MWA组)及肿瘤微波消融+瘤内接种DCs组(MWA+DC组).免疫组织化学法检测肿瘤组织内CD4+和CD8+T细胞的浸润,MTT法检测小鼠脾脏细胞对Hepa1-6的特异性杀伤活性,观测各组小鼠肿瘤生长情况.结果 免疫组织化学法检测显示MWA+DC组肿瘤组织内有大量的CD4+和CD8+T淋巴细胞浸润,显著高于其它组(P<0.05).MwA+DC组脾细胞对Hepa1-6细胞有特异性杀伤效能,在E/T=40和100时,MWA+DC组脾细胞对Hepa1-6细胞的特异性杀伤力显著高于对照组、DC组及MWA组(P<0.05).MWA+DC组小鼠肿瘤完全消退率显著高于其它各组(P<0.05).结论 MWA联合瘤内接种DCs可有效诱导机体产生特异性抗肝癌免疫,是预防MwA治疗后肝瘤复发的一种有效方法 .