前哨淋巴结定位检测在直肠癌外科中的应用研究

目的探讨定位直肠癌前哨淋巴结（SLN）方法以及临床应用价值。方法运用体内或体外注射亚甲蓝定位47例SLN,采用术中冰冻切片检查、HE染色病理检查、CK-20免疫组化染色（S—P法）检查检测SLN中转移癌。结果体内定位SLN成功32例,体外定位15例,成功14例。47例患者总淋巴结数目为849枚,平均为18．86枚／例,总SLN定位成功率为97．87％,平均SLN为1．87枚／例。常规病理检测SLN转移阴性26例,其中免疫组织化学方法检测CK-20发现微转移6例,上调23．08％（6／26）淋巴结转移阴性患者的病理分期。结论直肠癌体内、体外SIN定位均可获得成功;SLN CK-20免疫组化染色（S-P法）检查有助于提高早期直肠癌的病理分期准确率;SLN术中冰冻切片检查有助于术中指导切除范围。     

前哨淋巴结活检在结直肠癌根治术中的应用研究

目的探讨前哨淋巴结活检（SLNB）在结直肠癌根治术中临床应用的可行性及其价值。方法应用美蓝对67例结直肠癌患者行前哨淋巴结（SLN）定位活检,分体内、体外组,采用HE染色病理检查法、CK-20免疫组化染色（SP法）检测SLN中转移癌。结果共检出淋巴结660枚,其中SLN130枚,检出率19．7％。腹腔镜结直肠癌根治术和开腹结直肠癌根治术对SLN的检出差异无统计学意义（P=0．742）;体内、体外两种SLN的标记方法差异无统计学意义（P=0．564）;SP法检测SLN癌转移的敏感性明显高于HE染色,而假阴性率明显低于后者;肿瘤细胞在SLN的转移率明显高于区域淋巴结的转移率（P〈0．01）。结论结直肠癌根治术中体内、体外SLN定位方法均可以获得成功,均具有切实的可行性,与手术方式无关,并能够预测区域淋巴结的转移状况;通过SP法检查有助于明确结直肠癌的病理分期,有利于判断预后和个体化治疗方案的制定。     

结直肠癌前哨淋巴结体外标本定位及其微转移免疫组化研究

目的探讨结直肠癌前哨淋巴结（SLN）体外定位技术方法及其可行性、准确性和临床价值。方法选择2003年3月至2003年10月间中山大学肿瘤防治中心腹科住院行根治手术的结直肠癌患者60例,62个肿瘤（2例患者为多原发）,进行体外SLN定位。标本离体后尽早进行异硫蓝SLN定位,传统病理检查阴性的SLN进行细胞角蛋白免疫组化检查。结果62例肿瘤成功检出SLN的59例,检出率95．2％。59例患者总共获得并检测1114枚淋巴结,平均每人18．9（4～46）枚。其中SLN157枚（14．9％）,平均每人2．7（1～9）枚。SLN敏感性39．1％（9／23）,假阴性率23．7％（14／59）,准确率76．3％（45／59）。50例SLN阴性的中有12例（24％）细胞角蛋白免疫组化检测阳性。36例HE和细胞角蛋白免疫组化检查全阴性者中4例（11．1％）SLN发现微转移灶。14例仅非SLN阳性中8例SLN发现微转移灶。结论结直肠癌异硫蓝SLN体外定位活检技术是可行的,结合免疫组化检测微转移可以提高术后分期,可以提高送检淋巴结个数,结合免疫组化技术,可以减少淋巴结转移漏诊发生率。但该方法假阴性率较高,不能完全取代常规淋巴结病理检查。     

结直肠癌前哨淋巴结体外标记的研究

目的探讨结直肠癌前哨淋巴结（SEN）体外亚甲蓝定位活检方法的可行性，研究前哨淋巴结组织学状况能否用于预测区域淋巴结转移情况。方法将32例手术切除的结直肠癌标本纵行剪开，在癌肿四周注射亚甲蓝，2—5min后沿着蓝染的淋巴管追踪寻找首先蓝染的前哨淋巴结。将其切下后单独进行病理切片，检测有无癌转移，并与系膜淋巴结病理结果予以比较。结果有30例标本成功显示57枚SLN，平均每例标本显示1．9枚SLN。在SLN阳性的13例患者中。5例非SLN呈阳性，8例非SLN呈阴性；在17例SLN为阴性的标本中，15例非SLN呈阴性，仅2例非SLN呈阳性。统计本组患者SLN标记成功率为93．8％（30／32），准确率为93．3％（28／30），假阴性率为11．8％（2／17），特异性为100％（13／13）。结论结直肠癌标本前哨淋巴结体外亚甲蓝标记法可行，其组织学状况可较准确反映区域淋巴结群的癌转移情况。     

亚甲蓝前哨淋巴结定位在结直肠癌中的意义

目的探讨亚甲蓝存结直肠癌前哨淋巴结（sentinel lymph node,SLN）新鲜离体标本定位方法的可行性及临床意义。方法以1％业甲蓝对49例新鲜结直肠癌离体标本作SLN定位,标本染色后10分钟内先蓝染的第1—4个淋巴结视为SLN.所有标本均行常规石蜡切片．阴性的SI。N冉行多层切片,观察SLN对区域淋巴结转移的预测价值。结果49例病人中有47例检出SLN,检出率是95．92％,共获得SLN98枚,平均每例2．09枚（1～4枚）。SLN预测区域淋巴结转移的准确率为97．87％;特异性为95．83％;敏感性为94．45％;假阴性率为3．70％.结论以1％亚甲蓝作结直肠癌新鲜离体标本SLN定位简单、易行、可靠。SLN转移与否,可基本能反映区域淋巴结的转移状况。多层切片可降抵SLN的假阴性率。     

中低位直肠癌系膜内淋巴结微转移的临床病理学研究

目的 探讨中低位直肠癌全直肠系膜切除术（TME）后系膜内淋巴结微转移及其与临床病理因素的相关性。方法 31例中低位直肠癌患者接受了TME手术,应用脂肪清除技术获取全部淋巴结。将HE染色无转移的淋巴结标本进一步行抗CK抗体免疫组化染色,检测淋巴结内肿瘤微转移情况,并分析其与临床病理因素之间的相关性。结果 31例标本检出1007枚淋巴结。常规病理发现105枚淋巴结转移,直径为（5．5±3．1）mm;未见癌转移的淋巴结其直径为（2．8±1．5）mm,两组直径差异显著（P〈0．05）。在HE染色无转移的902枚淋巴结中,26枚发现微转移,其直径为（3．3±1．2）nq／n,显著小于常规病理有转移的淋巴结[（5．5±3．1）mm,P〈0．05],与常规病理无转移淋巴结直径差异不明显。有微转移的淋巴结其直径大部分〈5mm（88．5％）。31例患者中14例（38．7％）发现微转移;HE染色淋巴结无转移病例中微转移率为14．3％（2／14）。直肠系膜内淋巴结微转移与病理分期、血清癌胚抗原水平及肿瘤分化程度相关（P〈0．05）。结论 脂肪清除技术综合微转移的检测方法可使TME标本得到更多淋巴结,并能更加准确地判断淋巴结转移情况,有助于对直肠癌患者进行更加准确的临床分期并指导个体化治疗。     

结直肠癌前哨淋巴结微转移免疫组化检测的临床意义

目的探讨Cytokeratin-18(CK-18)免疫组织化学在结直肠癌前哨淋巴结转移检测中的应用价值。方法自2003年5月至2004年3月采用CK-18免疫组织化学方法检测99例SLN组织微转移癌。结果常规病理检测发现结直肠癌淋巴结转移45例,未转移54例。99例SLN常规病理癌转移阴性81例,其中CK-18免疫组织化学检测发现微转移31例。CK-18染色将15%(8/54)常规病理Dukes’A及B期患者肿瘤分期提升为Dukes’C期。CK-18免疫组织化学判定结直肠癌淋巴结转移的敏感性为98%,特异性为96%,准确度为97%。结论CK-18免疫组织化学检测结直肠癌SLN有助于提高Dukes’A及B期结直肠癌的病理分期。     

结直肠癌淋巴结微转移的临床研究

目的 探讨结直肠前哨淋巴结(SLN)的定位方法及检测淋巴结微转移的有效方法,并分析其临床意义.方法 对60例结直肠癌患者采用亚甲蓝染色法淋巴结示踪,寻找染色的SLN,切除后的SLN行HE染色和细胞角蛋白CK20免疫组化检测;并与前期直接行淋巴结清扫的60例患者对比.结果 亚甲蓝组中可识别SLN者54例(90.0%),高于前期直接清扫组的24例(40.0%)(P＜0.05);54例中行常规HE染色检出36例阳性,18例阴性.18例SLN阴性者行免疫组化检测,6例(33.3%)检出有微转移灶.结论 联合应用亚甲蓝和细胞角蛋白CK20进行结直肠癌SLN定位优于单用其中之一种方法;免疫组化是检测淋巴结微转移的敏感方法.     

362例结直肠癌手术切除标本淋巴结检出情况分析

分析结直肠癌患者手术切除标本淋巴结检出情况,探讨其与TNM分期、预后的关系。回顾性分析362例结直肠癌患者的临床资料和统计病理对淋巴结转移情况、随访资料。结果显示,有淋巴结转移（LNM）病例中,切除淋巴结数〉12枚者阳性率为73．1％,切除淋巴结数≤12枚者阳性率为58．0％（P〈0．05）。淋巴结转移率（rN）分期rN0、rN1、rN2 5年生存率分别为100％、84．4％和56．3％（P〈0．05）。结果表明,淋巴结切除数目影响LNM阳性率、rN和结直肠癌患者的预后判断。     

结直肠癌前哨淋巴结体外亚甲蓝染色的临床研究

目的探讨用体外亚甲蓝作为染色剂寻找前哨淋巴结(sentinel lymph node,SLN)的方法在结直肠癌SLN定位中的临床价值。方法将根治性切除大肠癌标本,在肿块四周浆膜下注射亚甲蓝后常规固定送病理检查。蓝染的淋巴结视为SLN。结果67例患者中找到SLN的有63例(91.3%),SLN镜下转移14例(22.22%)。结肠的SLN转移阳性的诊断价值与非SLN相比差异有统计学意义(P=0.0005),但在直肠癌中SLN转移阳性的诊断价值与非SLN相比差异无统计学意义(P=0.60),且SLN转移阳性在结肠癌病例中假阴性率为11.11%,而在直肠癌病例中则高达42.86%。结论体外亚甲蓝染色寻找SLN的方法在结肠癌中的应用是有效、可靠的,但在直肠癌中的应用其可靠性值得进一步的研究。     

前哨淋巴结活检在结直肠癌的研究与进展

前哨淋巴结是指原发肿瘤淋巴引流的第一级淋巴结.由于前哨淋巴结是肿瘤转移的第一站淋巴结,因此可以代表整个区域淋巴结状态.最近前哨淋巴结活检技术已经被广泛应用于结直肠癌.通过染色等方法找到前哨淋巴结后对其进行详细病理检查,发现微小转移,对于指导手术切除范围、准确病理分期以及指导术后治疗等方面具有重要意义.     

Ex vivo lymphatic mapping: a technique to improve pathologic staging in colorectal cancer

Sentinel lymph node (SLN) biopsy is widely used for solid tumors and has been proposed for use in staging colorectal cancer (CRC). Few studies have examined the ex vivo lymphatic mapping (EVLM) technique for staging CRC. We hypothesized that EVLM is technically feasible, sensitive, accurate, and improves the staging of CRC. After standard resection for colorectal cancer, 1 mL of isosulfan blue dye was injected circumferentially around the tumor. Blue-stained nodes were dissected separately and examined by hematoxylin and eosin (H&E) and immunohistochemical (IHC) stains. Routine pathologic evaluation was performed on all other harvested lymph nodes. Forty patients underwent 43 cancer resections with EVLM from July 2000 to December 2003. SLN were identified in 39 of 43 (91%) specimens. The mean number of SLN obtained was 1.9 (range, 0-5). Pathologic evaluation demonstrated nodal metastasis in 16 of 39 (39%) specimens. The SLN was tumor-positive in 9 of these 16 (56%) patients. The overall accuracy of EVLM was 82%. Two patients (9%) with H&E node negative disease were upstaged when found to have micrometastases by IHC staining. In conclusion, EVLM is technically possible in 90 per cent of patients with CRC. Although overall accuracy was high, the SLN status correlated poorly with the true nodal status of the CRC. However, EVLM improves pathologic staging in 9 per cent of patients and therefore may be of value in CRC.     

Sentinel lymph node evaluation is technically feasible and may improve staging in colorectal cancer

PURPOSE: The accurate nodal staging of colorectal cancer (CRC) is important to identify those patients who may benefit from adjuvant chemotherapy. Some have suggested that identification of sentinel lymph nodes (SLN) may improve staging in CRC. We sought to determine: the feasibility of identifying SLN in CRC utilizing isosulfan blue dye; the accuracy of the identified SLN in predicting the status of the remainder of the lymph nodes in CRC; and whether a more thorough evaluation of SLN with serial step sectioning and immunohistochemistry would more accurately stage patients with CRC. METHODS: A pilot trial was initiated at Walter Reed Army Medical Center (WRAMC), and 17 patients with masses on colonoscopy and subsequent tissue diagnosis of CRC were enrolled. Patients underwent standard surgical resection of their CRC with wedge of mesentery containing draining lymph nodes. Isosulfan blue dye was injected around the tumor subserosally/submucosally before dividing the mesenteric portion of the resection (n = 7) or ex vivo (n = 10). Sentinel lymph nodes were defined as all nodes staining blue and were dissected from the mesentery in the operating room. The SLN were sent separately for standard bivalving and hematoxylin and eosin staining (H&E) followed by serial step sectioning and immunohistochemistry (IHC) staining for cytokeratin. RESULTS: Seventeen patients (6 men, 11 women) were enrolled. The average preoperative carcinoembryonic antigen (CEA) was 5.9 (range, 1.2 to 18.9), and the average postoperative CEA was 2.8 (range, 0.7 to 9.1). One patient had a T1 tumor, 6 patients had T2 tumors, and 10 patients had T3 tumors on final pathology. Five cancers were well differentiated, 11 were moderately differentiated, and 1 was poorly differentiated. In all 17 cases, SLN were identified. A mean of 5.5 SLN was found per specimen (range, 2 to 11) with no difference noted between injection techniques (in vivo vs ex vivo). An additional 12 nonsentinel lymph nodes (range, 1 to 29) were identified per specimen. Ten patients had negative SLN and non-SLN. Seven patients were found to have positive SLN (3 by H&E, 2 by serial step sectioning, and 2 by IHC only). CONCLUSIONS: The isosulfan blue technique is technically feasible to allow identification of sentinel lymph nodes. In this study, no false-negative SLN occurred. A total of 7 patients had positive SLN; more importantly, 4 patients were upstaged as a result of serial step sectioning and immunohistochemistry staining. We hypothesize that this method may help pathologists find appropriate lymph nodes for more detailed analysis. As a result, patients may be more accurately staged and counseled for adjuvant chemotherapy, which has been shown to improve survival in node-positive CRC. Further studies should be undertaken to test these preliminary findings.     

Validation of Ex Vivo Lymphatic Mapping in Hematoxylin-Eosin Node-Negative Carcinoma of the Colon and Rectum

Background: Substantial evidence supports that detailed analysis of the regional lymphatics will identify previously unrecognized micrometastatic disease in colorectal cancer. In order to determine whether the sentinel lymph node(s) (SLNs) harvested by ex vivo lymphatic mapping in node-negative colorectal cancer (CRC) are the most likely node(s) to harbor micrometastatic disease, we examined all nodes in CRC specimens in an identical fashion.Methods: One hundred twenty-four specimens from patients with colorectal cancer were delivered to pathology in the fresh state and underwent ex vivo sentinel lymph node mapping. If negative by routine hematoxylin and eosin (H&E) analysis, the SLNs and non-SLNs were subjected to further analysis by level section H&E and immunohistochemical (IHC) analysis.Results: A mean of 30 nodes were harvested (range, 5–111). Fifty-one patients (41%) were found to be node-positive by routine H&E analysis. SLNs were identified in all but three specimens. A total of 2177 nodes were analyzed from the 66 H&E node-negative specimens (1883 non-SLNs and 294 SLNs). Overall, metastases were identified in 13 of 278 SLNs and in only 5 of 1829 non-SLNs (P < .001). Only 5 of 66 patients (7.5%) had evidence of metastatic disease in non-SLNs when the SLNs were negative. Thirteen apparently node-negative patients (19.3%) were upstaged by IHC analysis of the SLNs (P = .04).Conclusions: If the SLN is negative by both H&E and IHC analysis, the probability of finding metastases in a non-SLN is remote. If microstaging is demonstrated to be prognostically relevant, focused examination should be of the SLN(s).     

A novel lymphatic mapping technique to improve localization and staging of early colon cancer during laparoscopic colectomy

Encouraging results from our previous studies of sentinel lymph node (SLN) mapping in colorectal cancer (CRC) prompted investigation of its feasibility and accuracy during laparoscopic colectomy for early CRC. Between 1996 and 2000, 14 patients with clinically localized colorectal neoplasms underwent colonoscopic tattooing of the primary site and SLN mapping. In each case 0.5 to 1 cm3 of isosulfan blue dye was injected submucosally via the colonoscope. The blue-stained lymphatics were visualized through the laparoscope and followed to the SLN, which was marked with a clip, and laparoscopic colectomy was completed in the routine fashion. All lymph nodes were examined by hematoxylin and eosin (H&E) staining; in addition each SLN was subjected to focused examination by multisectioning and immunohistochemical staining using cytokeratin antibody. In all 14 patients the primary neoplasm and an SLN were identified laparoscopically. An average of 13.5 total lymph nodes and 1.7 SLNs per patient were identified. The SLN correctly reflected the tumor status of the nodal basin in 93 per cent of the cases. In four cases with unexpected lymphatic drainage, the extent of mesenteric resection was altered. In two cases (14%), nodal involvement was micrometastatic, confined to an SLN, and identified only by immunohistochemical staining. Lymphatic mapping caused no complications and added only 10 to 15 minutes to the overall operative time. Comparison of results in this group with results for a matched group of 14 patients undergoing SLN mapping during open colon resection showed that the laparoscopic technique had similar rates of accuracy and success. These preliminary findings indicate that colonoscopic/laparoscopic SLN mapping during laparoscopic colon resection is a feasible and technically simple means of identifying the primary colorectal neoplasm and its SLN. Focused pathologic examination of this node can upstage CRC and thereby may improve selection of patients for adjuvant chemotherapy.     

Focused examination of sentinel lymph nodes upstages early colorectal carcinoma

Approximately 30 per cent of patients with early colorectal carcinoma (CRC) develop systemic disease. A subgroup of these patients may harbor occult micrometastatic disease and might benefit from adjuvant chemotherapy. We investigated sentinel lymph node (SLN) mapping and focused pathologic examination of the SLN as a means of detecting nodal micrometastases. Between 1996 and 2000 SLN mapping was performed in 50 consecutive patients undergoing colectomy for CRC. All lymph nodes in the resection specimen were examined via routine hematoxylin and eosin (H&E) staining. In addition multiple sections of each SLN were examined via both H&E and cytokeratin immunohistochemistry. At least one SLN was identified in 47 patients (94%). In seven patients (14%) SLN mapping identified aberrant drainage that altered the planned resection. The SLN(s) correctly predicted nodal basin status in 44 of 47 (94%) cases; there were three falsely negative SLNs. Sixteen cases had positive SLNs by conventional H&E staining. An additional 10 (20%) cases were upstaged by a focused examination of the SLNs. Micrometastases were identified in three cases by H&E staining of multiple sections of the SLN and in seven only by cytokeratin immunohistochemistry. In nine cases the SLN was the only node containing tumor cells. In this study, SLN mapping demonstrated aberrant nodal drainage patterns that altered the surgical resection in patients with CRC. Focused examination of SLNs may detect micrometastases missed by conventional techniques and thereby identify patients who might benefit from adjuvant therapy.