Establishment of rapid molecular diagnoses of micrometastases of sentinel lymph node and its clinical application

我们的研究表明；1. 作为消化器官癌手术时扩张指标或淋巴结清扫减少指标的前哨淋巴结（SLN）的鉴别；3.肿瘤微转移的诊断；3 .外科手术中快速基因诊断方法的发展；SLN对预后影响的评价前哨淋巴结（SLN）的识别，作为消化器官癌手术时的扩张指标，或减少淋巴结清扫。常规病理诊断估计52例胃癌中30例淋巴结转移阴性的微转移情况。在评价7号淋巴结是否可作为SLN时，得出7号淋巴结可反映其他2组淋巴结转移情况的结果（假阴性率：16.7%，正确率：88.9%）。研究了苏木精染色、免疫组化、RT-PCR等方法对sln微转移的诊断。在常规HE染色评估的基础上，结合免疫组织化学和RT-PCR方法，成功地提高了SLN转移阳性病例的检出率（16），SLN的检出率：95→98%；[j] .中华医学会。2000)。外科手术中快速分子诊断方法的发展。采用常规RT-PCR方法获得基因诊断的诊断结果需要约8小时。我们成功地缩短了大约75分钟和时间，尽管目前通过改进RNA取样过程和引入实时PCR来获得诊断结果。当从新的RNA取样过程到实时PCR法的cDNA合成和检测技术正在建立时，估计获得诊断结果大约需要50分钟。评价SLN对预后的影响对28例7号淋巴结转移情况（RT-PCR诊断）进行分析，其中临床I期和II期的52例胃癌患者行根治性A切除。结果显示，7号淋巴结转移阳性患者预后较阴性患者差。少

Our study has revealed that ; 1. Identification of sentinel lymph node (SLN) used as index of expansion at time of operation of digestive organ cancer or reduction of lymph nodes dissection, 2. Diagnosis of micrometastases of SLN, 3. Development of rapid gene-diagnosis method while surgical operation, 4. Evaluation of influence of SLN affection on prognosis.1. Identification of sentinel lymph node (SLN) used as index of expansion at time of digestive organ cancer operation, or reduction of lymph nodes dissection.The conventional pathological diagnosis estimated the micrometastases situation of 30 cases that were lymph node metastases negative among 52 stomach cancer cases. The result in which #7 lymph node may reflect other 2 group lymph node may reflect other 2 group lymph nodes metastases situations in evaluation whether #7 lymph node may serve as SLN was obtaind (rate of false negative : 16.7 % and rate of accuracy : 88.9 %).2. Diagnosis of micrometastases of SLNThe hematoxylin-eosi … More n (HE) staining, the immunohistochemistry, and RT-PCR method have been examined. Then, it succeeded in raising the rate of detection of the cases, that is positive of metastases of SLN using together evaluation by the immunohistochemistry and RT-PCR method in addition to evaluation only using the conventional HE staining method (16 rate of SLN identification : 95→98 % ; Int. J. Oncol. 2000).3. Development of rapid molecular diagnosis method while surgical operation.It had required for obtaining a diagnostic result of the gene diagnosis using the conventional RT-PCR method for about 8 hours. We succeeded in shortening about 75 minutes and time, although a diagnostic result is obtained by an improvement of a RNA sampling process and introduction of real time PCR at present. When the technique from a new RNA sampling process to cDNA composition and delection by the real-time PCR method is now under establishment, it is presumed that it will be about 50 minutes obtaining a diagnostic result.4. Evaluation of influence SLN affects prognosisAnalysis about the metastases situation (it diagnoses in RT-PCR method) of #7 lymph node was performed about 28 cases in which were the clinical stage I and II performed curability A resection among 52 stomach cancer cases. It revealed that the cases of #7 lymph node metastases positive become poor prognosis compared with negative cases. Less