ⅠCDH1基因启动子区及内含子一单核苷酸多态性与膀胱移行细胞癌的相关研究 Ⅱ腹腔镜技术在肾脏功能保留和重建手术中的应用研究
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摘要
目的:检测中国湖北地区汉族非肿瘤人群CDH1基因近侧启动子、外显子一和内含子一单核苷酸多态性的类型和分布,并用生物信息学技术预测其功能。
材料和方法:以PCR-测序的方法对60例非肿瘤个体CDH1基因近侧启动子、外显子一和内含子一的单核苷酸多态性位点进行检测。用在线重复序列检测、转录因子结合位点分析和启动子分析软件对检测的单核苷酸多态性位点进行分析。
结果:在506 bp的测序长度中,共发现3个单核苷酸多态性,分布在启动子区2个,内含子一1个。启动子区2个单核苷酸多态性均不在重复序列内,-160C/A单核苷酸多态性可改变所在区域启动子活性,-73A/C单核苷酸多态性可使核转录结合因子发生改变。
结论:CDH1基因近侧启动子和内含子一共发现3个单核苷酸多态性,生物信息技术能提供有用信息,有助于指导进一步研究。
目的:探讨上皮钙粘素基因CDH1三个单核苷酸多态性位点和所组成的单体型与膀胱移行细胞癌易感性以及肿瘤病理特征的关系。
材料和方法:医院为基础的病例-对照研究。膀胱移行细胞癌病例130例,同一地区非肿瘤对照60例。以PCR-测序的方法对CDH1基因近侧启动子-160C/A、-73A/C和内含子一+178C/T三个单核苷酸多态性位点进行检测,比较不同基因型和等位基因型与个体膀胱移行细胞癌患病风险以及与肿瘤病理特征之间的关系;进一步构建并找出增加膀胱移行细胞癌易感性的单体型。
结果:-160C/A在膀胱移行细胞癌组和对照组分布有明显差异,肿瘤组携带A等位基因的比例高于对照组(P﹤0.01),并与肿瘤的分期成正相关(P﹤0.05)。-73A/C和178C/T在两组分布无明显差异(P﹥0.05)。三个单核苷酸多态性位点组成的基因型在两组分布不同,-160A/-73A/+178T单体型在膀胱移行细胞癌组(39.7%)的频率明显高于对照组(29.2%,P﹤0.001)。
结论:上皮钙粘素基因近侧调控序列-160处A等位基因频率与膀胱移行细胞癌的发生及其侵袭能力密切相关;AA基因型者膀胱移行细胞癌易感性比CC基因型者增加4.28倍;-160A/-73A/+178T单体型增加膀胱移行细胞癌的易感性。
目的:探讨后腹腔镜肾部分切除术治疗肾脏肿瘤的手术方法和临床效果。
方法:2002年6月至2004年2月间对32例为诊断为肾肿瘤的患者施行后腹腔镜保留肾单位手术(包括肿瘤剜除术和楔形切除术)。肿瘤平均直径2.8cm。主要使用超声刀进行肿瘤切除,结合使用超声刀、钛夹和腔内缝合技术止血。肾脏CT随访术后效果。
结果:32例手术全部在腹腔镜下完成,无中转开放者。剜除术平均手术时间70min,楔形切除术96min。剜除术平均术中出血量35.5ml,楔形切除术65.5ml。平均术后住院时间6.5d。术中术后无并发症。术后病检证实肾细胞癌21例,肾血管平滑肌脂肪瘤11例。在平均13个月的随访中,21例肾癌患者未见局部复发。
结论:后腹腔镜保留肾单位术治疗小的肾肿瘤安全可行,切除肿瘤精确彻底,创伤小,病人恢复快,值得临床有选择的推广使用。
目的:介绍后腹腔镜离断性肾盂成形术的手术方法并评价其临床效果。
方法:2000年5月至2004年9月间对50例确诊为肾盂输尿管连接部梗阻的患者施行腹腔镜离断性肾盂成形术。常规采用三孔后腹腔途径,气囊扩张法制备后腹腔空间。所有吻合采用腔内缝合技术。IVU和B超随访术后效果。
结果:50例手术全部在腹腔镜下完成,无中转开放者。平均手术时间81.6min(55～180min)。平均术中出血量12ml(5～50ml)。平均术后住院时间7.6d(6～12d)。术中无并发症。原发肾盂输尿管连接部狭窄44例,异位血管6例。2例出现吻合口漏,其中1例因梗阻复发接受开放手术。在平均22个月的随访中,49例患者肾盂输尿管连接部吻合口无狭窄,引流通畅。
结论:后腹腔镜离断性肾盂成形术是治疗肾盂输尿管连接部梗阻安全、有效及微创的方法,值得临床推广使用。
目的:通过与开放离断性肾盂成形术的效果比较,评价后腹腔镜离断性肾盂成形
术的临床价值。方法:回顾分析后腹腔镜离断性肾盂成形术56例(A组)及开放离断性肾盂成形术40例(B组)的临床资料,就两组手术时间、术中出血量、术后肠道功能恢复时间、术后止疼药用量、术后住院天数、并发症和手术成功率等指标进行比较。根据数据类型选用X2检验、成组T检验或Mann-Whitey U检验。
结果:A组在手术时间、术中出血量、术后恢复饮食时间、术后止疼药用量、手术切口长度和术后住院天数方面优于B组,差别有极显著性意义(P<0.001);并发症和成功率与B组相当,差异无统计学意义(P=0.058)。
结论:后腹腔镜离断性肾盂成形术是治疗肾盂输尿管连接部梗阻的一种微创、安全、有效的方法。存在较明显的学习曲线,熟练后可在较短时间完成手术。
Objectives: The single nucleotide polymorphisms in the proximum promoter, exon 1 and intron 1 regions of E-cadherin gene were detected; and their potential function were analysised by bioinformatics software. Materials and methods: We screened the proximum promoter, exon 1 and intron 1 regions of CDH1 for single nucleotide polymorphism in 60 Chinese using direct sequences. The allele frequencty of the single nucleotide polymorphism was calculated. The potential function of the single nucleotide polymorphism was analysised by three bioinformatics softwares.
Results: Three single nucleotide polymorphisms were observed. Two of them (-160 C/A and -73 A/C) were in proximum promotor region and one (+178 C/T) in intron 1 region. The -160C/A and +178T/C polymorphisms are high frequency SNPs and the frequencies are 48.3% and 28.3% respectively. The frequenciy of -73A/C polymorphisms is 13.3%. Bioinformatics analysis showed that the -73C/A polymorphism is responsible for binding of several nuclear transcription factors and -160 C/A polymorphism locates in one pomoter region.
Conclusions: Three single nucleotide polymorphisms were observed. The Bioinformatics Analysis can provide useful information to facilitate the further study.
Part Two: Association of SNPs and Haplotypes in the Proximum Promotor Region and Intron 1 of CDH1 Gene with Transitional Cell Carcinoma of Urinary Bladder
Objectives: To investigate whether the three single nucleotide polymorphisms and haplotypes in the proximum promoter region and intron 1 of CDH1 are associated with transitional cell carcinoma of urinary bladder.
Materials and methods: A hospital-based case-control study was performed on 130 patients with TCCB (male 96 and female 34, age form 29 to 84 years, mean 58.1±14.7) and 60 normal controls (male 44 and female 16, age form 28 to 81 years, mean 58.1±14. 7). Genomic DNA was extracted from blood samples of the subjects. Genotypes were determined using direct sequences. The allele, genotype and haplotypes frequencies of the three single nucleotide polymorphisms and their association with transitional cell carcinoma of urinary bladder were analysised. Appropriate tests were selected for statistical analysis.
Results: We observed a significantly higher frequency of the -160A allele in TCCB patients (76%) compared to that in the control group (64%) (P﹤0.05). The A allele frequencies at the same position were also significantly higher in invasive TCCB than in superficial carcinoma(P﹤0.05); however there was no statistical difference in the A allele frequency between high pathological grade and low pathological grade of TCCB (P >0.05). When compared TCCB patients to the control group, there was no statistical difference for the allele frequency in -73A/C and +178C/T. The distribution of haplotypes is different in both groups. There is a a significantly higher frequency of -160A/-73A/+178T haplotype in TCCB patients (39.7%) compared to that in the control group (29.2%) (P﹤0.001).
Conclusions: The -160C/A single nucleotide polymorphism in the proximum regulatory region of E-cadherin gene is associated with TCCB; the -160A/-73A/+178T haplotype increases the susceptibility with TCCB, and this A-A-T haplotype might prove a useful marker.
Purpose: To evaluate the feasibility and clinical efficacy of retroperitoneal laparoscopic nephron-sparing surgery for the renal tumors.
Materials and Methods: Between June 2002 and February 2004, 11 cases of renal benign tumor and 21 cases of renal malignant tumor underwent enucleation of the tumor and wedge resection of the tumor through retroperitoneal laparoscopy respectively. Tumor resection and hemostasis were achieved mainly by Harmonic Scalpel. Follow-up studies were performed with an evaluation for renal spiral computed tomography. Results: All procedures were technically successful. Mean operating time was 70 min for enucleation and 96 min for wedge resection. Mean estimated blood loss was 35 ml for enucleation and 65.5 ml for wedge resection. Mean hospital stay after operation was 6.5 days. No intraoperational complications occurred. Pathological examination confirmed renal cell carcinoma (RCC) in 21 patients and angiomyolipoma in 11.
Pathological stage was pT1a in 21 RCC patients. All resected tumor specimens had negative surgical margins for cancer. No local recurrence or trocar site metastasis was observed during a follow-up of mean 13 months.
Conclusions: Our results indicate that retroperitoneal laparoscopic nephron-sparing surgery represents a feasible option for patients with localized renal tumors. This procedure could offer precise and complete tumor excision while minimizing morbidity, improving cosmesis and shortening convalescence.
Purpose: We report our techniques and experience of retroperitoneal laparoscopic dismembered pyeloplasty for ureteropelvic junction (UPJ) obstruction.
Materials and Methods: From May 2000 to September 2004, 50 patients with UPJ obstruction underwent laparoscopic dismembered pyeloplasty. A three-port, balloon-dissecting, retroperitoneal approach was used. All anastomoses were completed with free hand intracorporeally suture techniques. Follow-up studies were performed with IVU and renal ultrasonography.
Results: All operations were completed laparoscopically and no open conversions were required. The mean operating time was 81.6 min (ranging from 55 to 180 min). The mean blood loss was 12 ml (ranging from 5 to 50 ml) and the mean postoperative hospitalization day was 7.6 days (ranging from 6 to 12 days). No intraoperative complications occurred. Aberrant artery vessel and primary stricture as cause of UPJ obstruction was noted in 6 and 44 patients respectively. Anastomotic leakage occurred in 2 patients and one of them received open surgery for recurrent UPJ obstruction later. Radiographic assessment by IVU showed good results in 49 cases with a mean follow-up of 22 months.
Conclusions: Our experience with retroperitoneal laparoscopic dismembered pyeloplasty demonstrates that this technique is an effective treatment for UPJ obstruction and can be accomplished reasonably quickly.
Purpose: To evaluate the clinical value of retroperitoneal laparoscopic dismembered pyeloplasty for ureteropelvic junction obstruction as compared with open surgery. Materials and Methods: The clinical data of 56 patients who underwent retroperitoneal laparoscopic dismembered pyeloplasty were retrospectively compared with those of 40 patients who underwent open dismembered pyeloplasty through a retroperitoneal flank approach. Student t-test, Pearson Chi-square test and Mann-Whitney rank sum test were applied for statistical analysis as appropriate. Results: Patient's demographic data were similar between the two groups. In the laparoscopic group, operative time (80 versus 120minutes), estimated blood loss (10 versus 150mL), recovery of intestinal function (1 versus 2days), analgesic requirements (Diclofenac Sodium Suppository) (75 versus 150mg), incision length (3.5versus 21cm), and postoperative hospital stay (7 versus 9days) were better than those in the open group (P<0.001 for all). No intraoperative complications occurred in either group. Incidence of postoperative complications (2/56, 3.6% versus 3/40, 7.5%, P=0.729) and success rate (55/56, 98.2% versus 39/40, 97.5%, P=0.058) were equivalent in the two groups.
Conclusions: Retroperitoneal laparoscopic dismembered pyeloplasty is a minimally invasive, safe and effective therapy for ureteropelvic junction obstruction with low morbidity, shorter convalescence and excellent outcomes and can be accomplished reasonably quickly in experienced hands.
材料和方法:以PCR-测序的方法对60例非肿瘤个体CDH1基因近侧启动子、外显子一和内含子一的单核苷酸多态性位点进行检测。用在线重复序列检测、转录因子结合位点分析和启动子分析软件对检测的单核苷酸多态性位点进行分析。
结果:在506 bp的测序长度中,共发现3个单核苷酸多态性,分布在启动子区2个,内含子一1个。启动子区2个单核苷酸多态性均不在重复序列内,-160C/A单核苷酸多态性可改变所在区域启动子活性,-73A/C单核苷酸多态性可使核转录结合因子发生改变。
结论:CDH1基因近侧启动子和内含子一共发现3个单核苷酸多态性,生物信息技术能提供有用信息,有助于指导进一步研究。
目的:探讨上皮钙粘素基因CDH1三个单核苷酸多态性位点和所组成的单体型与膀胱移行细胞癌易感性以及肿瘤病理特征的关系。
材料和方法:医院为基础的病例-对照研究。膀胱移行细胞癌病例130例,同一地区非肿瘤对照60例。以PCR-测序的方法对CDH1基因近侧启动子-160C/A、-73A/C和内含子一+178C/T三个单核苷酸多态性位点进行检测,比较不同基因型和等位基因型与个体膀胱移行细胞癌患病风险以及与肿瘤病理特征之间的关系;进一步构建并找出增加膀胱移行细胞癌易感性的单体型。
结果:-160C/A在膀胱移行细胞癌组和对照组分布有明显差异,肿瘤组携带A等位基因的比例高于对照组(P﹤0.01),并与肿瘤的分期成正相关(P﹤0.05)。-73A/C和178C/T在两组分布无明显差异(P﹥0.05)。三个单核苷酸多态性位点组成的基因型在两组分布不同,-160A/-73A/+178T单体型在膀胱移行细胞癌组(39.7%)的频率明显高于对照组(29.2%,P﹤0.001)。
结论:上皮钙粘素基因近侧调控序列-160处A等位基因频率与膀胱移行细胞癌的发生及其侵袭能力密切相关;AA基因型者膀胱移行细胞癌易感性比CC基因型者增加4.28倍;-160A/-73A/+178T单体型增加膀胱移行细胞癌的易感性。
目的:探讨后腹腔镜肾部分切除术治疗肾脏肿瘤的手术方法和临床效果。
方法:2002年6月至2004年2月间对32例为诊断为肾肿瘤的患者施行后腹腔镜保留肾单位手术(包括肿瘤剜除术和楔形切除术)。肿瘤平均直径2.8cm。主要使用超声刀进行肿瘤切除,结合使用超声刀、钛夹和腔内缝合技术止血。肾脏CT随访术后效果。
结果:32例手术全部在腹腔镜下完成,无中转开放者。剜除术平均手术时间70min,楔形切除术96min。剜除术平均术中出血量35.5ml,楔形切除术65.5ml。平均术后住院时间6.5d。术中术后无并发症。术后病检证实肾细胞癌21例,肾血管平滑肌脂肪瘤11例。在平均13个月的随访中,21例肾癌患者未见局部复发。
结论:后腹腔镜保留肾单位术治疗小的肾肿瘤安全可行,切除肿瘤精确彻底,创伤小,病人恢复快,值得临床有选择的推广使用。
目的:介绍后腹腔镜离断性肾盂成形术的手术方法并评价其临床效果。
方法:2000年5月至2004年9月间对50例确诊为肾盂输尿管连接部梗阻的患者施行腹腔镜离断性肾盂成形术。常规采用三孔后腹腔途径,气囊扩张法制备后腹腔空间。所有吻合采用腔内缝合技术。IVU和B超随访术后效果。
结果:50例手术全部在腹腔镜下完成,无中转开放者。平均手术时间81.6min(55～180min)。平均术中出血量12ml(5～50ml)。平均术后住院时间7.6d(6～12d)。术中无并发症。原发肾盂输尿管连接部狭窄44例,异位血管6例。2例出现吻合口漏,其中1例因梗阻复发接受开放手术。在平均22个月的随访中,49例患者肾盂输尿管连接部吻合口无狭窄,引流通畅。
结论:后腹腔镜离断性肾盂成形术是治疗肾盂输尿管连接部梗阻安全、有效及微创的方法,值得临床推广使用。
目的:通过与开放离断性肾盂成形术的效果比较,评价后腹腔镜离断性肾盂成形
术的临床价值。方法:回顾分析后腹腔镜离断性肾盂成形术56例(A组)及开放离断性肾盂成形术40例(B组)的临床资料,就两组手术时间、术中出血量、术后肠道功能恢复时间、术后止疼药用量、术后住院天数、并发症和手术成功率等指标进行比较。根据数据类型选用X2检验、成组T检验或Mann-Whitey U检验。
结果:A组在手术时间、术中出血量、术后恢复饮食时间、术后止疼药用量、手术切口长度和术后住院天数方面优于B组,差别有极显著性意义(P<0.001);并发症和成功率与B组相当,差异无统计学意义(P=0.058)。
结论:后腹腔镜离断性肾盂成形术是治疗肾盂输尿管连接部梗阻的一种微创、安全、有效的方法。存在较明显的学习曲线,熟练后可在较短时间完成手术。
Objectives: The single nucleotide polymorphisms in the proximum promoter, exon 1 and intron 1 regions of E-cadherin gene were detected; and their potential function were analysised by bioinformatics software. Materials and methods: We screened the proximum promoter, exon 1 and intron 1 regions of CDH1 for single nucleotide polymorphism in 60 Chinese using direct sequences. The allele frequencty of the single nucleotide polymorphism was calculated. The potential function of the single nucleotide polymorphism was analysised by three bioinformatics softwares.
Results: Three single nucleotide polymorphisms were observed. Two of them (-160 C/A and -73 A/C) were in proximum promotor region and one (+178 C/T) in intron 1 region. The -160C/A and +178T/C polymorphisms are high frequency SNPs and the frequencies are 48.3% and 28.3% respectively. The frequenciy of -73A/C polymorphisms is 13.3%. Bioinformatics analysis showed that the -73C/A polymorphism is responsible for binding of several nuclear transcription factors and -160 C/A polymorphism locates in one pomoter region.
Conclusions: Three single nucleotide polymorphisms were observed. The Bioinformatics Analysis can provide useful information to facilitate the further study.
Part Two: Association of SNPs and Haplotypes in the Proximum Promotor Region and Intron 1 of CDH1 Gene with Transitional Cell Carcinoma of Urinary Bladder
Objectives: To investigate whether the three single nucleotide polymorphisms and haplotypes in the proximum promoter region and intron 1 of CDH1 are associated with transitional cell carcinoma of urinary bladder.
Materials and methods: A hospital-based case-control study was performed on 130 patients with TCCB (male 96 and female 34, age form 29 to 84 years, mean 58.1±14.7) and 60 normal controls (male 44 and female 16, age form 28 to 81 years, mean 58.1±14. 7). Genomic DNA was extracted from blood samples of the subjects. Genotypes were determined using direct sequences. The allele, genotype and haplotypes frequencies of the three single nucleotide polymorphisms and their association with transitional cell carcinoma of urinary bladder were analysised. Appropriate tests were selected for statistical analysis.
Results: We observed a significantly higher frequency of the -160A allele in TCCB patients (76%) compared to that in the control group (64%) (P﹤0.05). The A allele frequencies at the same position were also significantly higher in invasive TCCB than in superficial carcinoma(P﹤0.05); however there was no statistical difference in the A allele frequency between high pathological grade and low pathological grade of TCCB (P >0.05). When compared TCCB patients to the control group, there was no statistical difference for the allele frequency in -73A/C and +178C/T. The distribution of haplotypes is different in both groups. There is a a significantly higher frequency of -160A/-73A/+178T haplotype in TCCB patients (39.7%) compared to that in the control group (29.2%) (P﹤0.001).
Conclusions: The -160C/A single nucleotide polymorphism in the proximum regulatory region of E-cadherin gene is associated with TCCB; the -160A/-73A/+178T haplotype increases the susceptibility with TCCB, and this A-A-T haplotype might prove a useful marker.
Purpose: To evaluate the feasibility and clinical efficacy of retroperitoneal laparoscopic nephron-sparing surgery for the renal tumors.
Materials and Methods: Between June 2002 and February 2004, 11 cases of renal benign tumor and 21 cases of renal malignant tumor underwent enucleation of the tumor and wedge resection of the tumor through retroperitoneal laparoscopy respectively. Tumor resection and hemostasis were achieved mainly by Harmonic Scalpel. Follow-up studies were performed with an evaluation for renal spiral computed tomography. Results: All procedures were technically successful. Mean operating time was 70 min for enucleation and 96 min for wedge resection. Mean estimated blood loss was 35 ml for enucleation and 65.5 ml for wedge resection. Mean hospital stay after operation was 6.5 days. No intraoperational complications occurred. Pathological examination confirmed renal cell carcinoma (RCC) in 21 patients and angiomyolipoma in 11.
Pathological stage was pT1a in 21 RCC patients. All resected tumor specimens had negative surgical margins for cancer. No local recurrence or trocar site metastasis was observed during a follow-up of mean 13 months.
Conclusions: Our results indicate that retroperitoneal laparoscopic nephron-sparing surgery represents a feasible option for patients with localized renal tumors. This procedure could offer precise and complete tumor excision while minimizing morbidity, improving cosmesis and shortening convalescence.
Purpose: We report our techniques and experience of retroperitoneal laparoscopic dismembered pyeloplasty for ureteropelvic junction (UPJ) obstruction.
Materials and Methods: From May 2000 to September 2004, 50 patients with UPJ obstruction underwent laparoscopic dismembered pyeloplasty. A three-port, balloon-dissecting, retroperitoneal approach was used. All anastomoses were completed with free hand intracorporeally suture techniques. Follow-up studies were performed with IVU and renal ultrasonography.
Results: All operations were completed laparoscopically and no open conversions were required. The mean operating time was 81.6 min (ranging from 55 to 180 min). The mean blood loss was 12 ml (ranging from 5 to 50 ml) and the mean postoperative hospitalization day was 7.6 days (ranging from 6 to 12 days). No intraoperative complications occurred. Aberrant artery vessel and primary stricture as cause of UPJ obstruction was noted in 6 and 44 patients respectively. Anastomotic leakage occurred in 2 patients and one of them received open surgery for recurrent UPJ obstruction later. Radiographic assessment by IVU showed good results in 49 cases with a mean follow-up of 22 months.
Conclusions: Our experience with retroperitoneal laparoscopic dismembered pyeloplasty demonstrates that this technique is an effective treatment for UPJ obstruction and can be accomplished reasonably quickly.
Purpose: To evaluate the clinical value of retroperitoneal laparoscopic dismembered pyeloplasty for ureteropelvic junction obstruction as compared with open surgery. Materials and Methods: The clinical data of 56 patients who underwent retroperitoneal laparoscopic dismembered pyeloplasty were retrospectively compared with those of 40 patients who underwent open dismembered pyeloplasty through a retroperitoneal flank approach. Student t-test, Pearson Chi-square test and Mann-Whitney rank sum test were applied for statistical analysis as appropriate. Results: Patient's demographic data were similar between the two groups. In the laparoscopic group, operative time (80 versus 120minutes), estimated blood loss (10 versus 150mL), recovery of intestinal function (1 versus 2days), analgesic requirements (Diclofenac Sodium Suppository) (75 versus 150mg), incision length (3.5versus 21cm), and postoperative hospital stay (7 versus 9days) were better than those in the open group (P<0.001 for all). No intraoperative complications occurred in either group. Incidence of postoperative complications (2/56, 3.6% versus 3/40, 7.5%, P=0.729) and success rate (55/56, 98.2% versus 39/40, 97.5%, P=0.058) were equivalent in the two groups.
Conclusions: Retroperitoneal laparoscopic dismembered pyeloplasty is a minimally invasive, safe and effective therapy for ureteropelvic junction obstruction with low morbidity, shorter convalescence and excellent outcomes and can be accomplished reasonably quickly in experienced hands.
引文
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