大肠癌“湿热瘀毒”证候病机与相关肿瘤标志物的临床研究
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摘要
目的:本实验旨在深入研究古典医籍及现代中医文献的基础上,并结合前期临床研究,阐明“湿热瘀毒”是大肠癌的中医证候病机的关键,对大肠癌重新进行中医辨证分型,制定大肠癌的中医“湿热瘀毒”证型及其辨证标准,并对大肠癌“湿热瘀毒”证手术病人的肿瘤原发灶、肿瘤周边淋巴结和癌旁正常上皮组织进行相关肿瘤标志物(CEA、CA242、CA-199和CK20)的联合检测,进一步确定“湿热瘀毒”证候大肠癌相关肿瘤标志物的特殊表达,以指导临床的诊断治疗和判断预后。
方法:
1理论研究
本研究是在深入研究古典医籍及现代中医文献的基础上,以中医病因病机理论为依据,收集、整理、提炼,结合前期临床研究,提出“湿热瘀毒”是大肠癌中医病因病机关键的科学假说,并制定以“湿热瘀毒”证候为主的大肠癌中医辨证分型。
2临床研究
①临床研究以大肠癌患者为研究对象,根据中医理论及结合临床,通过病人的临床症状体征等,在重新进行大肠癌中医辨证分型的基础上,制定中医“湿热瘀毒”证候大肠癌的辨证标准。
②通过免疫组织化学方法分别对50例大肠癌“湿热瘀毒”证手术病人和50例正常人为对照组,对肿瘤原发灶、肿瘤周边淋巴结、癌旁正常上皮组织进行相关肿瘤标志物(CEA、CA242、CA-199)的联合检测,进一步确定肿瘤的特殊表达,肿瘤的分期,并将其与临床资料和预后相联系,指导临床及时开展术后化疗及监测。
③通过免疫组织化学方法和RT-PCR方法,分别对大肠癌“湿热瘀毒”证的手术病人不同组织的CK20mRNA及蛋白表达影响的实验研究,根据其特殊表达情况,指导临床及时开展术后化疗及监测。
结果:
1理论研究结果
首次提出“湿热瘀毒”是大肠癌的中医证候病机的关键,并阐述大肠癌形成“湿热瘀毒”证候病机的理论根据;在大肠癌辨证论治分型中首先提出“湿热瘀毒”证型。
2临床研究结果
(1)提出大肠癌辨证论治可分为以下四个证型:①湿热瘀毒证、②脾肾阳虚证、③肝肾阴虚证、④气血两虚证。其中以“湿热瘀毒”为主要证型,并制定中医“湿热瘀毒”证候大肠癌的辨证标准。本研究对2006年6月至2007年9月期间收治的102例大肠癌病人,进行中医辨证分型。其中属于湿热瘀毒证的有63例;属于脾肾阳虚证的有11例;属于肝肾阴虚证的有13例;属于气血两虚证的有15例。湿热瘀毒证所占比例为61.8%。
(2)大肠癌“湿热瘀毒”证的手术病人不同组织CEA蛋白表达影响的实验研究:大肠癌“湿热瘀毒”证患者肿瘤原发灶的CEA相对其他各组有明显的表达,CEA对大肠癌有较高的敏感性,但缺少特异性,不能作为肿瘤的筛选指标,而是用于肿瘤患者的监测和疗效的判断。
(3)大肠癌“湿热瘀毒”证的手术病人不同组织CA242蛋白表达影响的实验研究:大肠癌“湿热瘀毒”证患者,CA242在大肠癌组织中呈高度表达,而在良性组织中很少表达。而CA242在肿瘤周边淋巴结组的阳性表达,它的增高预示着肿瘤的微转移。
(4)大肠癌“湿热瘀毒”证的手术病人不同组织CA19-9蛋白表达影响的实验研究:大肠癌“湿热瘀毒”证患者,CA19-9在大肠癌组织中呈高度表达,对大肠癌有较高的敏感性;而在良性组织中很少表达。研究发现平行联合检测CEA、CA242和CAl9-9可明显提高肿瘤检测的阳性率,利用它们的阳性互补性,优于任一单项肿瘤标志物。
(5)大肠癌“湿热瘀毒”证的手术病人不同组织中CK20及CK20mRNA表达影响的实验研究:CK20在良性组织中很少表达,但在大肠癌组织中呈高度表达。大肠癌“湿热瘀毒”证患者,通过RT- PCR法测定大肠癌组织淋巴结中CK20mRNA表达,通过其表达程度判断微转移比传统的病理形态学更敏感,对大肠癌患者能做出准确的预后判断。
结论:
1研究证实“湿热瘀毒”是大肠癌的中医证候病机的关键。
2制定了大肠癌的中医“湿热瘀毒”证型及其辨证标准。
3阐明大肠癌“湿热瘀毒”证患者肿瘤原发灶的CEA相对其他各组有明显的表达,CEA对大肠癌有较高的敏感性,但缺少特异性。
4阐明大肠癌“湿热瘀毒”证患者,CA242在大肠癌组织中呈高度表达,CA242在良性组织中很少表达。
5大肠癌“湿热瘀毒”证患者,CA19-9在大肠癌组织中呈高度表达,而在良性组织中很少表达,对大肠癌有较高的敏感性。
6 CK20在良性组织中很少表达,但在大肠癌组织中呈高度表达,通过RT- PCR法测定大肠癌组织淋巴结的微转移比传统的病理形态学更敏感,对大肠癌患者能做出准确的预后判断。
7分析“湿热瘀毒”证大肠癌的证候病机,有助于中医临床的辨证论治。采用CEA、CA19-9、CA242、CK20四项肿瘤标志物联合检测,有助于提高肿瘤标记物诊断的敏感性和特异性,提高肿瘤的阳性诊断率,可更好的判断肿瘤的分期和预后,有助于提高对大肠癌的诊断准确性。
Objective:
The experiment is based on the deep study on classical medical books and modern TCM literature, combining with clinical research early days. It is first believed that“damp-heat stagnancy toxin”is the key of TCM etiology and pathogenesis of colorectal carcinoma. It also brings forward the clinical presentations syndrome of“damp-heat stagnancy toxin”of colorectal carcinoma and constitutes its diagnostic criteria in TCM, which refreshes the differential syndrome of TCM. Third, it has a united test of relevant tumor markers(CEA、CA242、CA19-9 and CK20) about“damp-heat stagnancy toxin”syndrome operating patients on tumor idiopathic place、tumor peripheral lymphnodes and normal epithelium tissues beside tumor,so that it further identifies the special expression of relevant tumor markers on“damp-heat stagnancy toxin”syndrome of colorectal carcinoma,which can guide clinic and judge prognosis
Method:
1 Theoretical Research
Basing on the deep study on classical medical books and modern TCM literature and according to the theory of TCM collateral diseases, we puts forward the scientific hypothesis that“damp-heat stagnancy toxin”is the key of etiology and pathogenesis in TCM and constitutes that“damp-heat stagnancy toxin”syndrome is the main clinical presentations of TCM differential typing on colorectal carcinoma by collection、arrangement and abstraction, also in combination with forward clinical research.
2 Clinical Research
①The clinical research constitutes diagnostic criteria of TCM“damp-heat stagnancy toxin”syndrome of colorectal carcinoma, which is based on studying colorectal carcinoma patients, according to the basic theory of TCM , combining with clinical presentations of the patients, and refreshing differential typing of TCM on colorectal carcinoma.
②Through the method of immunohistochemistry ,we have an united test of relevant tumor markers(CEA、CA242、CA19-9 and CK20) about“damp-heat stagnancy toxin”syndrome operating patients on tumor aidiopathic place、tumor peripheral lymphnodes and normal epithelium tissues beside tumor, so as to further identify tumors special expression, tumors stage, also in combination with clinical materials and prognosis to guide clinic and develop anonym and monitoring after operation.
③Using the method of immunohistochemistry and RT-PCR ,the experiment respectively studies the expression of protein CK20 and CK20mRNA in different tissue, which is from“damp-heat stagnancy toxin”syndrome operating patients of colorectal carcinoma. Through its special expression, it guides clinic and develop anonym and monitoring after operation.
Results
1 Results of theoretical research
It first puts forward and elaborates the theoretical frame and foundation that how“damp-heat stagnancy toxin”clinical presentations、pathogenesis of colorectal carcinoma forms. It thinks that“damp-heat stagnancy toxin”syndrome is the key of pathogenesis of colorectal carcinoma in TCM. It first brings forward“damp-heat stagnancy toxin”syndrome in differential typing of colorectal carcinoma.
2 results of clinical research
①We first put forward that the differential treatment is divided into four syndromes: 1) damp-heat stagnancy toxin syndrome 2)Yang deficiency of the spleen and kidney syndrome 3) Yin deficiency of liver and kidney syndrome 4) deficiency of qi and blood syndrome. Among them,“damp-heat stagnancy toxin”syndrome is the main type. We also constitute the diagnostic criteria of“damp-heat stagnancy toxin”syndrome of colorectal carcinoma in TCM. We have a differential typing in TCM among 102 colorectal carcinoma patients in hospital from 2006.6 to 2007.9 .Among them, damp-heat stagnancy toxin: 63; Yang deficiency of the spleen and kidney: 11; Yin deficiency of liver and kidney: 13; deficiency of qi and blood: 15; damp-heat stagnancy toxin syndrome is 61.8%.
②The experimental research on the expression of protein CEA in different tissues of“damp-heat stagnancy toxin”syndrome operating patients suffered from colorectal carcinoma .It has obvious expression that CEA of tumor idiopathic place from“damp-heat stagnancy toxin”syndrome patients of colorectal carcinoma compared with other groups. CEA is more sensitive for colorectal carcinoma. Because CEA has lower specific, it can’t be the select norm. So it is only used to monitor the patient of tumor and judge the therapeutic effectiveness
③The experimental research on the expression of protein CA242 in different tissues of“damp-heat stagnancy toxin”syndrome operating patients suffered from colorectal carcinoma For“damp-heat stagnancy toxin”syndrome patients suffered from colorectal carcinoma, it highly expresses that protein CA242 in colorectal carcinoma tissues and seldom expresses in benign tissues. Furthermore,CA242 is the positive expression in peripheral lymphonotes of tumor. Its heightening indicates the micromitastasis of tumor.
④The experimental research on the expression of protein CA19-9 in different tissues of“damp-heat stagnancy toxin”syndrome operating patients suffered from colorectal carcinoma.For“damp-heat stagnancy toxin”syndrome patients of colorectal carcinoma, protein CA19-9 highly expresses in colorectal carcinoma tissues. It is more sensitive for colorectal carcinoma and seldom expresses in benign tissues. The research shows that the positive rate could be obviously increased by parallelly and unitedly testing CEA、CA242 and CA19-9. It is better than doing only one test, using their positive complementarities.
⑤The experimental research on the expression of CK20 and CK20mRNA in different tissues of“damp-heat stagnancy toxin”syndrome operating patients suffered from colorectal carcinoma.CK20 seldom expresses in benign tissues but highly expresses in colorectal carcinoma tissues. For“damp-heat stagnancy toxin”syndrome patients of colorectal carcinoma,it is more sensitive than traditional pathomorphology test that we use the method of RT-PCR to test micromitastasis of lymphonote of colorectal carcinoma tissues, so we can give a exact prognostic judgment.
Conclusions:
①The research testifies that“damp-heat stagnancy toxin”is the key of etiology and pathogenesis of colorectal carcinoma in TCM.
②Establishing the clinical presentations syndrome of“damp-heat stagnancy toxin”of colorectal carcinoma in TCM and its diagnostic criteria.
③Clarifying the obvious expression of CEA in tumor idiopathic place from“damp-heat stagnancy toxin”syndrome patients of colorectal carcinoma compared with other groups. CEA is more sensitive for colorectal carcinoma but has lower specific
④Clarifying that CA242 is highly expresses in colorectal carcinoma tissues and seldom expresses in benign tissues from“damp-heat stagnancy toxin”syndrome patients of colorectal carcinoma.
⑤For“damp-heat stagnancy toxin”syndrome patients of colorectal carcinoma , protein CA19-9 highly expresses in colorectal carcinoma tissue and seldom expresses in benign tissues. CA19-9 is more sensitive for colorectal carcinoma.
⑥CK20 seldom expresses in benign tissues but highly expresses in colorectal carcinoma tissues. It is more sensitive than traditional pathomorphology test that we use RT-PCR to test micromitastasis of lymphnodes of colorectal carcinoma tissues, so we can give an exact prognostic judgment.
⑦In this research, we found that the analysis on the cause and pathogenesis of“damp-heat stagnancy toxin”syndrome about colorectal carcinoma will contribute to the clinical differentiation in TCM. Sensibility and specific of tumor markers and positive diagnostic rate could be obviously increased by unitedly testing four kinds of tumor markers CEA、CA242 and CA19-9. It is better at judging neoplasm staging and prognosis and helpful for increasing diagnostic accuracy of colorectal carcinoma.
方法:
1理论研究
本研究是在深入研究古典医籍及现代中医文献的基础上,以中医病因病机理论为依据,收集、整理、提炼,结合前期临床研究,提出“湿热瘀毒”是大肠癌中医病因病机关键的科学假说,并制定以“湿热瘀毒”证候为主的大肠癌中医辨证分型。
2临床研究
①临床研究以大肠癌患者为研究对象,根据中医理论及结合临床,通过病人的临床症状体征等,在重新进行大肠癌中医辨证分型的基础上,制定中医“湿热瘀毒”证候大肠癌的辨证标准。
②通过免疫组织化学方法分别对50例大肠癌“湿热瘀毒”证手术病人和50例正常人为对照组,对肿瘤原发灶、肿瘤周边淋巴结、癌旁正常上皮组织进行相关肿瘤标志物(CEA、CA242、CA-199)的联合检测,进一步确定肿瘤的特殊表达,肿瘤的分期,并将其与临床资料和预后相联系,指导临床及时开展术后化疗及监测。
③通过免疫组织化学方法和RT-PCR方法,分别对大肠癌“湿热瘀毒”证的手术病人不同组织的CK20mRNA及蛋白表达影响的实验研究,根据其特殊表达情况,指导临床及时开展术后化疗及监测。
结果:
1理论研究结果
首次提出“湿热瘀毒”是大肠癌的中医证候病机的关键,并阐述大肠癌形成“湿热瘀毒”证候病机的理论根据;在大肠癌辨证论治分型中首先提出“湿热瘀毒”证型。
2临床研究结果
(1)提出大肠癌辨证论治可分为以下四个证型:①湿热瘀毒证、②脾肾阳虚证、③肝肾阴虚证、④气血两虚证。其中以“湿热瘀毒”为主要证型,并制定中医“湿热瘀毒”证候大肠癌的辨证标准。本研究对2006年6月至2007年9月期间收治的102例大肠癌病人,进行中医辨证分型。其中属于湿热瘀毒证的有63例;属于脾肾阳虚证的有11例;属于肝肾阴虚证的有13例;属于气血两虚证的有15例。湿热瘀毒证所占比例为61.8%。
(2)大肠癌“湿热瘀毒”证的手术病人不同组织CEA蛋白表达影响的实验研究:大肠癌“湿热瘀毒”证患者肿瘤原发灶的CEA相对其他各组有明显的表达,CEA对大肠癌有较高的敏感性,但缺少特异性,不能作为肿瘤的筛选指标,而是用于肿瘤患者的监测和疗效的判断。
(3)大肠癌“湿热瘀毒”证的手术病人不同组织CA242蛋白表达影响的实验研究:大肠癌“湿热瘀毒”证患者,CA242在大肠癌组织中呈高度表达,而在良性组织中很少表达。而CA242在肿瘤周边淋巴结组的阳性表达,它的增高预示着肿瘤的微转移。
(4)大肠癌“湿热瘀毒”证的手术病人不同组织CA19-9蛋白表达影响的实验研究:大肠癌“湿热瘀毒”证患者,CA19-9在大肠癌组织中呈高度表达,对大肠癌有较高的敏感性;而在良性组织中很少表达。研究发现平行联合检测CEA、CA242和CAl9-9可明显提高肿瘤检测的阳性率,利用它们的阳性互补性,优于任一单项肿瘤标志物。
(5)大肠癌“湿热瘀毒”证的手术病人不同组织中CK20及CK20mRNA表达影响的实验研究:CK20在良性组织中很少表达,但在大肠癌组织中呈高度表达。大肠癌“湿热瘀毒”证患者,通过RT- PCR法测定大肠癌组织淋巴结中CK20mRNA表达,通过其表达程度判断微转移比传统的病理形态学更敏感,对大肠癌患者能做出准确的预后判断。
结论:
1研究证实“湿热瘀毒”是大肠癌的中医证候病机的关键。
2制定了大肠癌的中医“湿热瘀毒”证型及其辨证标准。
3阐明大肠癌“湿热瘀毒”证患者肿瘤原发灶的CEA相对其他各组有明显的表达,CEA对大肠癌有较高的敏感性,但缺少特异性。
4阐明大肠癌“湿热瘀毒”证患者,CA242在大肠癌组织中呈高度表达,CA242在良性组织中很少表达。
5大肠癌“湿热瘀毒”证患者,CA19-9在大肠癌组织中呈高度表达,而在良性组织中很少表达,对大肠癌有较高的敏感性。
6 CK20在良性组织中很少表达,但在大肠癌组织中呈高度表达,通过RT- PCR法测定大肠癌组织淋巴结的微转移比传统的病理形态学更敏感,对大肠癌患者能做出准确的预后判断。
7分析“湿热瘀毒”证大肠癌的证候病机,有助于中医临床的辨证论治。采用CEA、CA19-9、CA242、CK20四项肿瘤标志物联合检测,有助于提高肿瘤标记物诊断的敏感性和特异性,提高肿瘤的阳性诊断率,可更好的判断肿瘤的分期和预后,有助于提高对大肠癌的诊断准确性。
Objective:
The experiment is based on the deep study on classical medical books and modern TCM literature, combining with clinical research early days. It is first believed that“damp-heat stagnancy toxin”is the key of TCM etiology and pathogenesis of colorectal carcinoma. It also brings forward the clinical presentations syndrome of“damp-heat stagnancy toxin”of colorectal carcinoma and constitutes its diagnostic criteria in TCM, which refreshes the differential syndrome of TCM. Third, it has a united test of relevant tumor markers(CEA、CA242、CA19-9 and CK20) about“damp-heat stagnancy toxin”syndrome operating patients on tumor idiopathic place、tumor peripheral lymphnodes and normal epithelium tissues beside tumor,so that it further identifies the special expression of relevant tumor markers on“damp-heat stagnancy toxin”syndrome of colorectal carcinoma,which can guide clinic and judge prognosis
Method:
1 Theoretical Research
Basing on the deep study on classical medical books and modern TCM literature and according to the theory of TCM collateral diseases, we puts forward the scientific hypothesis that“damp-heat stagnancy toxin”is the key of etiology and pathogenesis in TCM and constitutes that“damp-heat stagnancy toxin”syndrome is the main clinical presentations of TCM differential typing on colorectal carcinoma by collection、arrangement and abstraction, also in combination with forward clinical research.
2 Clinical Research
①The clinical research constitutes diagnostic criteria of TCM“damp-heat stagnancy toxin”syndrome of colorectal carcinoma, which is based on studying colorectal carcinoma patients, according to the basic theory of TCM , combining with clinical presentations of the patients, and refreshing differential typing of TCM on colorectal carcinoma.
②Through the method of immunohistochemistry ,we have an united test of relevant tumor markers(CEA、CA242、CA19-9 and CK20) about“damp-heat stagnancy toxin”syndrome operating patients on tumor aidiopathic place、tumor peripheral lymphnodes and normal epithelium tissues beside tumor, so as to further identify tumors special expression, tumors stage, also in combination with clinical materials and prognosis to guide clinic and develop anonym and monitoring after operation.
③Using the method of immunohistochemistry and RT-PCR ,the experiment respectively studies the expression of protein CK20 and CK20mRNA in different tissue, which is from“damp-heat stagnancy toxin”syndrome operating patients of colorectal carcinoma. Through its special expression, it guides clinic and develop anonym and monitoring after operation.
Results
1 Results of theoretical research
It first puts forward and elaborates the theoretical frame and foundation that how“damp-heat stagnancy toxin”clinical presentations、pathogenesis of colorectal carcinoma forms. It thinks that“damp-heat stagnancy toxin”syndrome is the key of pathogenesis of colorectal carcinoma in TCM. It first brings forward“damp-heat stagnancy toxin”syndrome in differential typing of colorectal carcinoma.
2 results of clinical research
①We first put forward that the differential treatment is divided into four syndromes: 1) damp-heat stagnancy toxin syndrome 2)Yang deficiency of the spleen and kidney syndrome 3) Yin deficiency of liver and kidney syndrome 4) deficiency of qi and blood syndrome. Among them,“damp-heat stagnancy toxin”syndrome is the main type. We also constitute the diagnostic criteria of“damp-heat stagnancy toxin”syndrome of colorectal carcinoma in TCM. We have a differential typing in TCM among 102 colorectal carcinoma patients in hospital from 2006.6 to 2007.9 .Among them, damp-heat stagnancy toxin: 63; Yang deficiency of the spleen and kidney: 11; Yin deficiency of liver and kidney: 13; deficiency of qi and blood: 15; damp-heat stagnancy toxin syndrome is 61.8%.
②The experimental research on the expression of protein CEA in different tissues of“damp-heat stagnancy toxin”syndrome operating patients suffered from colorectal carcinoma .It has obvious expression that CEA of tumor idiopathic place from“damp-heat stagnancy toxin”syndrome patients of colorectal carcinoma compared with other groups. CEA is more sensitive for colorectal carcinoma. Because CEA has lower specific, it can’t be the select norm. So it is only used to monitor the patient of tumor and judge the therapeutic effectiveness
③The experimental research on the expression of protein CA242 in different tissues of“damp-heat stagnancy toxin”syndrome operating patients suffered from colorectal carcinoma For“damp-heat stagnancy toxin”syndrome patients suffered from colorectal carcinoma, it highly expresses that protein CA242 in colorectal carcinoma tissues and seldom expresses in benign tissues. Furthermore,CA242 is the positive expression in peripheral lymphonotes of tumor. Its heightening indicates the micromitastasis of tumor.
④The experimental research on the expression of protein CA19-9 in different tissues of“damp-heat stagnancy toxin”syndrome operating patients suffered from colorectal carcinoma.For“damp-heat stagnancy toxin”syndrome patients of colorectal carcinoma, protein CA19-9 highly expresses in colorectal carcinoma tissues. It is more sensitive for colorectal carcinoma and seldom expresses in benign tissues. The research shows that the positive rate could be obviously increased by parallelly and unitedly testing CEA、CA242 and CA19-9. It is better than doing only one test, using their positive complementarities.
⑤The experimental research on the expression of CK20 and CK20mRNA in different tissues of“damp-heat stagnancy toxin”syndrome operating patients suffered from colorectal carcinoma.CK20 seldom expresses in benign tissues but highly expresses in colorectal carcinoma tissues. For“damp-heat stagnancy toxin”syndrome patients of colorectal carcinoma,it is more sensitive than traditional pathomorphology test that we use the method of RT-PCR to test micromitastasis of lymphonote of colorectal carcinoma tissues, so we can give a exact prognostic judgment.
Conclusions:
①The research testifies that“damp-heat stagnancy toxin”is the key of etiology and pathogenesis of colorectal carcinoma in TCM.
②Establishing the clinical presentations syndrome of“damp-heat stagnancy toxin”of colorectal carcinoma in TCM and its diagnostic criteria.
③Clarifying the obvious expression of CEA in tumor idiopathic place from“damp-heat stagnancy toxin”syndrome patients of colorectal carcinoma compared with other groups. CEA is more sensitive for colorectal carcinoma but has lower specific
④Clarifying that CA242 is highly expresses in colorectal carcinoma tissues and seldom expresses in benign tissues from“damp-heat stagnancy toxin”syndrome patients of colorectal carcinoma.
⑤For“damp-heat stagnancy toxin”syndrome patients of colorectal carcinoma , protein CA19-9 highly expresses in colorectal carcinoma tissue and seldom expresses in benign tissues. CA19-9 is more sensitive for colorectal carcinoma.
⑥CK20 seldom expresses in benign tissues but highly expresses in colorectal carcinoma tissues. It is more sensitive than traditional pathomorphology test that we use RT-PCR to test micromitastasis of lymphnodes of colorectal carcinoma tissues, so we can give an exact prognostic judgment.
⑦In this research, we found that the analysis on the cause and pathogenesis of“damp-heat stagnancy toxin”syndrome about colorectal carcinoma will contribute to the clinical differentiation in TCM. Sensibility and specific of tumor markers and positive diagnostic rate could be obviously increased by unitedly testing four kinds of tumor markers CEA、CA242 and CA19-9. It is better at judging neoplasm staging and prognosis and helpful for increasing diagnostic accuracy of colorectal carcinoma.
引文
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