应用改良AP—PCR技术进行食管癌相关基因分析

目的 应用改良的随机引物聚合酶链反应技术，分离配对的食管癌组织和非癌食管组织中差异基因片断，并进行克隆，测序和序列分析。方法 在来自食管癌高发区的２２例配对的食管癌组织和非癌食管组织中，应用ＡＰ－ＰＣＲ技术测检了差异基因片段。结果 ２２例配对食管癌组织和非癌食管组织中，６例癌组织中有差异的随机扩增片断，而相应的非癌组织缺如。     

人食管癌相关基因cDNA片段的克隆与初步鉴定

目的 分离人原发性食管组织中瓣的相关基因,揭示食管癌的易感性与癌变原理,方法 用高效,灵敏的ｍＲＮＡ差异显示技术,以２例正常食管上皮,１例癌旁上皮,２例高癌家族的食管癌组织互为对照,通过其对其因表达的比较,找出差异条带,进行ＲＴ－ＰＣＲ鉴定和ＤＮＡ序列分析,结果 （１）在实验中,分离,鉴定了１８个差异片段,其中包括正常组织不表达的突变食管基因（ｍｕｔａｔｅｄｅｓｏｐｈａｇｅａｌｇｅｎｅ,ＭＥＧ）５     

食管癌组织中MTS1/p16基因的缺失

目的 研究ＭＴＳ１／ｐ１６基因在人食管组织及相应癌旁组织中的变化。方法 采用Ｓｏｕｔｈｅｒｎ杂交和和方法检测了６０例人食管组织标本（包括３０例食管癌和３０例相应癌旁上皮组织）中Ｐ１６基因的缺失情况。结果 在３０例癌旁上皮组织中未检测到Ｐ１６基因缺失,而在３０例食管癌组织中,Ｓｏｕｔｈｅｒｎ检测有７例标本Ｐ１６基因杂交阴怀,ＰＣＲ扩增证实其中有５例标本Ｐ１６基因缺失;Ｐ１６基因在食管癌组织中的缺失频     

MAL基因在人食管癌中表达显著下调

研究ＭＡＬ基因在人食管癌中的表达下调状况。方法采用Ｎｏｒｔｈｅｒｎ杂交和ＲＴ－ＰＣＲ方法检测了ＭＡＬ基因在４１对配对的食管癌组织和相应癌旁食管粘膜中的表达;应用ＰＣＲ和ＲＴ－ＰＣＲ方法分析了ＭＡＬ基因在３个食管癌细胞系ＥＣ１０９、ＥＣ８７１２和ＥＣ９７０６中的存在及其表达。     

食管癌组织中抑癌基因APC,MCC突变的研究

应用聚合酶链反应（ＰＣＲ）扩增与直接测序方法，分析抑癌基因ＡＰＣ、ＭＣＣ在食管癌中的变化，应用ＰＣＲ扩增，发现１／１０的食管癌组织有ＡＰＣ基因第１１外显子缺失，１／１０的食管癌组织有ＭＣＣ基因第１２外显子缺失，并发现１例食管癌旁癌组织有ＭＣＣ基因基因第１２个外显子缺失。ＰＣＲ直接测序发现：２／１０的食管癌标本有ＡＰＣ基因第１１外显子突变，２／７的食管癌组织有ＭＣＣ基因第１２外显子突变。以上研究证实     

食管癌和贲门癌组织中P53基因突变的研究

应用核酸杂交、ＰＣＲ扩增、直接测序等分子生物学技术，分析了林县原发性食管癌和贲门癌及其癌旁组织中抑癌基因Ｐ５３人子结构改变。用Ｓｏｕｔｈｅｍ杂交法分析的３５例食管鳞癌和２７例组织中，有１例Ｐ５３基因缺失，１例重排结构性改变，占７．４％。上述食管鳞癌标本，用ＰＣＲ直接测序，发现５０％９１５／３０）的食管鳞癌有Ｐ５３基因的的５－９外显子和相应内含突变。４５％（５／１１）食管癌旁组织中有Ｐ５３基因突变。     

食管癌组织端粒酶活性的研究

目的：检测食管癌及周围非癌食管粘膜活性、了解食管癌发生可能的分子生物学基础。方法：应用ＴＲＡＰ法对３３例食管及１８例癌周正常食管粘膜进行端粒酶活性测定。结果：食管癌组织２８例（８４．８％）表达阳性,癌周组织２例（１１．１％）表达阳性。端粒酶活性表达与食管的性别,民族,肿瘤部位,分化程度,有无淋巴结转移等似乎无关。结论：端粒酶活化与食管癌发生有关,并可为食管肿瘤早期诊断与治疗提供可靠的指标与线索     

食管癌端粒酶活性检测的意义

目的 探讨食管癌组织中端粒酶活性检测的临床意义。方法 采用聚合酶链反应－酶联免疫吸附法（ＰＣＲ－ＥＬＩＳＡ）检测了４３例食管癌组织及相应癌旁组织,８例食管良性疾病组织端粒酶活性表达。结果 ４３例食管癌组织中３６例端粒酶活性阳性表达,阳性率为８３．７％,而癌旁组织４例阳性,阳性率为９．３％,８例良性疾病食管组织端粒酶表达均为阴性。食管癌端粒酶检测阳性率明显高于癌旁组织及良性疾病食管组织。食管癌组织端     

人食管癌细胞系中缺失的新基因片段在食管癌发生中的可能意义

目的寻找食管癌的抑癌基因,揭示食管癌的遗传易感性和癌变原理。方法采用Ｓｏｕ－ｔｈｅｒｎ杂交和ＰＣＲ方法,检测食管癌细胞系中缺失的三个ＤＮＡ片段（１２Ｈ２、３３Ｂ２和３３Ｂ３）,在２０例食管癌组织和相应的癌旁组织及三个高危家族成员外周血中的缺失情况。结果在癌和癌旁组织标本中,１２Ｈ２的缺失频率最高,癌组织为６１．１％,癌旁组织为２２．２％,淋巴结转移阳性率为４５．４％;３３Ｂ２分别为３０．０％、１０．０％和５０．０％;３３Ｂ３分别为２０．０％、１０．０％和２５．０％。１２Ｈ２和３３Ｂ３在三个高危家族中未发现种系改变和先证者的体细胞纯合缺失。３３Ｂ２在一个家系的先证者中存在体细胞纯合缺失,但未发现种系改变。结论１２Ｈ２、３３Ｂ２和３３Ｂ３可能为新的食管癌候选抑癌基因,且与食管癌的进展和转移相关。     

食管癌组织中金属硫蛋白3基因甲基化的地区性差异

目的:探讨食管癌高发区河北省中南部及广东省潮汕地区和食管癌非高发区的食管癌及正常组织中金属硫蛋白3 (metallothionein-3, MT-3)基因CpG岛的甲基化情况及其临床意义.方法:选取正常人血液标本10例,胎儿食管组织标本10例,非食管癌高发区正常人食管黏膜标本20例,河北省中南部食管癌高发区和潮汕地区食管癌新鲜手术标本及切缘正常组织各30例;应用甲基化特异性聚合酶链反应 (methylation-specific PCR, MSP)检测其MT-3基因CpG岛的甲基化情况,并分析其与临床病理资料之间的关系.结果: 高发区食管癌患者的切缘正常组织有20例(33.3%)存在MT-3基因甲基化,高于非高发区正常人食管黏膜(P=0.013);食管癌组织中有49例(81.7%)甲基化,明显高于相应肿瘤切缘正常组织(P<0.001),但2个高发区之间比较差异无统计学意义(P=0.739).结论:食管癌患者的正常食管黏膜及癌组织中均广泛存在MT-3基因的甲基化,后天因素刺激可能是导致MT-3基因甲基化的重要原因.     

食管癌、贲门癌术后心律失常的原因分析

目的 探讨食管癌、贲门癌术后心律失常的原因.方法 对我科收治的600例食管癌、贲门癌患者临床资料进行回顾性分析.结果 600例食管癌、贲门癌患者术后共发生心律失常136例.不同年龄、术前心电图表现、肿瘤部位的患者术后心律失常的发生率:＜60岁者和≥60岁者分别为10.4％、25.0％;术前心电图检查正常者和异常者分别为6.3％、87.6％;食管癌、贲门癌患者分别为24.5％、14.4％,各组比较差异均有统计学意义(P均＜0.05).结论 高龄、术前心电图异常、手术方式、术后并发症与术后心律失常的发生密切相关.     

Molecular Biology of Esophageal Cancer

There have been many new developments in our understanding of esophageal carcinoma biologyover the past several years.Information regarding both of the major forms of this disease,adenocarcinomaand squamous cell carcinoma,has accumulated in conjunction with data on precursor conditions such asBarrett's esophagus.Interesting and promising findings have included overexpression of proto-oncogenes,loss of heterozygosity at multiple chromosomal loci,tumor suppressor gene inactivation,epigenetic silenc-ing by DNA methylation,and mutations and deletions involving the tumor suppressor gene p53.Importantcancer pathways,the cyclin kinase inhibitor cascade and the DNA mismatch repair process,implicatedin the genesis of multiple tumor types have also been inculpated in esophageal carcinogenesis.Alterationsin the p16 and p15 cyclin kinase inhibitors including point mutations and homozygous deletions havebeen reported in primary esophageal tumors.Further developments in the field of molecular carcinogen-esis of esophageal malignancies promise to yield improvements in prevention,early detection,prognosticcategorization,and perhaps gene-based therapy of this deadly disease.     

食管癌的分子生物学

There have been many new developments in our understanding of esophageal carcinoma biology over the past several years. Information regarding both of the major forms of this disease, adenocarcinoma and squamous cell carcinoma, has accumulated in conjunction with data on precursor conditions such as Barrett‘s esophagus. Interesting and promising findings have included overexpression of proto-oncogenes,loss of heterozygosity at multiple chromosomal loci, tumor suppressor gene inactivation, epigenetic silencing by DNA methylation, and mutations and deletions involving the tumor suppressor gene p53. Important cancer pathways, the cyclin kinase inhibitor cascade and the DNA mismatch repair process, implicated in the genesis of multiple tumor types have also been inculpated in esophageal carcinogenesis. Alterations in the p16 and p15 cyclin kinase inhibitors including point mutations and homozygous deletions have been reported in primary esophageal tumors. Further developments in the field of molecular carcinogenesis of esophageal malignancies promise to yield improvements in prevention, early detection, prognostic categorization, and perhaps gene-based therapy of this deadly disease.     

p53 Gene Mutations in Esophageal Cancer Detected by Polymerase Chain Reaction Single-strand Conformation Polymorphism Analysis

Mutations of the p53 gene play an important role in the development of common human malignancies. We investigated mutations of this gene in 26 surgical specimens of esophageal cancer using the polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) analysis. The results were correlated with histological findings, DNA ploidy and the short-term relapse of the disease. PCR-SSCP analysis detected mutations of the p53 gene in 10 tumors (38%), eight in exons 5–6 and two in exons 7–8. A higher incidence of lymph node metastasis, poorly differentiated tumor, DNA aneuploidy and short-term relapse of the disease was observed in cases with p53 gene mutations, although the findings were not statistically significant.     

食管贲门癌手术死亡原因分析

目的　分析食管贲门癌手术后死亡原因 ,探讨有效的预防措施。方法　对我院近 10年来 3 3例食管贲门癌手术死亡病例的临床资料进行分析总结。结果　本组手术死亡率为 1 1% ,死亡原因占前三位的分别是心脑血管疾病、吻合口瘘、呼吸系统疾病 ;手术死亡率与手术范围、病期及年龄明显相关。结论　严格掌握手术适应证 ,选择合理的手术范围及术式 ,积极仔细的围手术期处理 ,正确处理术后并发症 ,对降低食管贲门癌的手术死亡率有重要意义     

食管癌的新辅助治疗

Because the conflicting data currently available from the performed randomized trials it is very difficult to provide strict guidelines for the treatment of patients with locoregional advanced esophageal cancers. Surgery however, remains the standard of care for potentially resectable disease. Preoperative chemotherapy is still controversial with two large randomized trials resulting in two different conclusions regarding the survival benefit. Preoperative chemoradiation is also controversial since only one randomized trial showed a clear survival benefit however, the patients treated with surgery alone in this trial had an unusually poor outcome. And the study by Urba et al was not powered enough to show a clear survival benefit for patients treated with neoadjuvant chemoradiation. The results of three metaanalysis of these randomized studies show lower rate of resection, higher rate of R0-resection, more often postoperative mortality and better prognosis for patients with neoadjuvant radiochemotherapy. As a consequence one may consider offering neoadjuvant chemotherapy or neoadjuvant radiochemotherapy to patients with locallyadvanced disease under the premise that patients have a good performance status and understand the controversies about this therapeutic option. Larger trials with sufficient power to clearly detect survival benefits for patients treated with neoadjuvant chemotherapy or radiochemotherapy are necessary before this therapeutic option will be the standard of care.     

食管和贲门癌组织中肿瘤抑制基因maspin的表达

 采用免疫组织化学（ABC）法检测食管癌高发区55例食管和贲门癌组织中肿瘤抑制基因maspin的表达。结果表明,食管癌组织中maspin免疫阳性表达率为10%（3/30）,免疫阳性反应主要位于癌组织的细胞浆中;贲门癌组织中maspin免疫阳性表达率为24%（6/25）,免疫阳性反应主要位于癌组织的细胞核中。提示：肿瘤抑制基因maspin可能在食管和贲门癌变过程中起一定的作用。     

Neoadjuvant Treatment for Esophageal Cancer

Because the conflicting data currently available from the performed randomized trials it is verydifficult to provide strict guidelines for the treatment of patients with locoregional advanced esophagealcancers.Surgery however,remains the standard of care for potentially resectable disease.Preoperativechemotherapy is still controversial with two large randomized trials resulting in two different conclusionsregarding the survival benefit.Preoperative chemoradiation is also controversial since only one randomizedtrial showed a clear survival benefit however,the patients treated with surgery alone in this trial had anunusually poor outcome.And the study by Urba et al was not powered enough to show a clear survivalbenefit for patients treated with neoadjuvant chemoradiation.The results of three metaanalysis of theserandomized studies show lower rate of resection,higher rate of RO-resection,more often postoperativemortality and better prognosis for patients with neoadjuvant radiochemotherapy.As a consequence one mayconsider offering neoadjuvant chemotherapy or neoadjuvant radiochemotherapy to patients with locally-advanced disease under the premise that patients have a good performance status and understand thecontroversies about this therapeutic option.Larger trials with sufficient power to clearly detect survivalbenefits for patients treated with neoadjuvant chemotherapy or radiochemotherapy are necessary beforethis therapeutic option will be the standard of care.     

宫颈癌和食管癌组织中人乳头状瘤病毒基因型的检测

[目的]探讨宫颈癌及食管癌组织中人乳头状瘤病毒（HPV）感染及其基因型分布的相关性。 [方法]采用基因芯片方法分别检测200例宫颈癌和140例食管癌的HPV基因型,计算HPV感染率,分析HPV基因型与癌肿类型的关系。 [结果]200例宫颈癌的HPV阳性率为94．00％（188／200）,共检测到9种HPV基因型,其中主要基因型为HPV16（74．00％）和HPV18（16．00％）,HPV双重感染的阳性率达12．00％（24／2001。169例宫颈鳞癌的HPV阳性率为95．86％,显著高于腺癌的80．00％（x2=9．73,P〈0．01）,但HPV16、18和HPV双重感染在鳞、腺癌中的阳性率无显著性差异。在140例食管癌组织中,未检测到任何基因型别的HPV。 [结论]宫颈癌组织的HPV感染型别多样,HPV16、18为最常见类型,HPV感染更常见于宫颈鳞癌。但HPV各型别的分布与癌肿类型无关．HPV似乎与食管癌的发生无关。     

青年人食管贲门癌临床特点及误诊原因分析

目的　分析青年人食管贲门癌的特点及误诊原因。方法　收集了 3 5岁以下青年人食管贲门癌的病例资料。结果　青年人呈现发病率低、易误诊、恶性程度高、病变进展快、转移发生早、手术切除率低、5年生存率低等特点。结论　提高青年患者生存率的关键是早期发现、早期诊断、早期治疗。