48例直肠癌前哨淋巴结微转移检测

[目的]探讨前哨淋巴结（SLN）微转移检测在提高直肠癌病理分期及预测区域淋巴结转移中的价值。[方法]2003年12月至2005年12月行根治性手术的48例直肠癌患者采用同位素标记的方法进行SLN定位检查,SLN行常规病理检查,并进一步应用免疫组织化学（IHC）和逆转录聚合酶链式反应（PCR）方法进行微转移的检测。[结果]48例患者共检出SLN108枚,占淋巴结总数的12.22%（108/884）。经HE/IHC/PCR检查,48例患者中,SLN转移者共38例,无转移者10例,SLN预测区域淋巴结转移状态的准确率为79.17%（38/48）。常规病理前仅SLN阳性2例,检测微转移后仅SLN阳性12例,10例患者病理分期由Dukes’B期上升到C期。[结论]应用CK-20蛋白或其mRNA表达检测同位素示踪法所检出的直肠癌SLN微转移情况,具有较高的预测直肠癌区域淋巴结转移状态的价值,并有助于提高直肠癌病理分期的准确性。     

巢式RT-PCR检测乳腺癌前哨淋巴结微转移的研究

目的：探讨乳腺癌前哨淋巴站（SLN）定位和腋淋巴结微转移检测的临床意义,方法：对20例乳腺癌患者术中肿块周围注射美蓝定位前哨淋巴 结,用巢式RT－PCR法检测腋淋巴结中Mammaglobin mRNA的表达,结果：SLN定位成功率为85．0％（17／20）,SLN与非SLN组微转移的检出率具显著差异（P＜0．01）。在常规病检阴性的淋巴结中,巢式RT－PCR法微转移的检出率为15．6％（17／109）,结论：巢式RT－PCR法较常规病理检查更为敏感,通过SLN定位和巢式RT－PCR的联合使用,可明显提高乳腺癌腋淋巴结微转移的检出效率。     

乳腺癌前哨淋巴结CK19 mRNA和蛋白的表达

目的 探讨CK19微转移分子在乳腺癌前哨淋巴结(SLN)的表达情况及意义.方法 采用免疫组化S-P法及原位杂交技术检测66例SLN中CK19微转移分子的表达.同时与常规病检法比较其检测敏感性.并比较转移组、微转移组、无转移组患者的临床资料.结果 66例SLN在常规病理检查阴性38例淋巴结中6例表达CK19蛋白15.8%(6/38),8例21.1%(8/38)表达CK19mRNA.原位杂交法与常规病检法转移的检出率相比较,差异有统计学意义(P＜0.05).CK19蛋白和mRNA表达之间差异无统计学意义(P＞0.05).同时乳腺癌转移组与微转移组患者在肿物大小与淋巴管浸润上有相似性,而同无转移组差异有统计学意义(P＜0.05).结论 免疫组化法和原位杂交法较常规病理检查更为敏感.通过免疫组化法和原位杂交法的联合使用,可明显提高乳腺癌SLN微转移的检出率,同时也证明原位杂交法是可靠的,比免疫组化更为敏感的技术.SLN微转移有可能作为肿瘤预后的指标.     

结直肠癌前哨淋巴结体外定位检测的临床研究

目的:探讨用体外亚甲蓝作为染色剂寻找前哨淋巴结(sentinellymph node,SLN)的方法在结直肠癌SLN定位中的可行性及临床价值。方法:将行标准根治性切除的结直肠癌标本离体后,在肿块四周黏膜下注射亚甲蓝后追踪辨认。蓝染的淋巴结视为SLN,未蓝染的淋巴结被视为NSLN。SLN中无癌细胞转移者常规行细胞角蛋白(CK、AE1/AE3)免疫组化检查,CK阳性者视为有微转移病例。结果:82例患者成功标记SLN(96.47%),准确度为90.24%。通过CK检测,16例SLN阴性患者发现微转移,总转移率提升了18.30%,TNM分期得以提升的患者达18.82%。HE染色下SLN的假阴性率为23.17%,结直肠癌的假阴性率分别为6.38%和45.71%,有统计学意义（P=0.001）。结论:前哨淋巴结活检（SLNB）对结肠癌区域淋巴结转移情况的判断更有临床价值,但在直肠癌方面的应用值得进一步探讨。     

前哨淋巴结活检术在结直肠癌中的应用研究

目的:探讨结直肠癌中前哨淋巴结定位活检术的适用性。方法:20例结直肠癌患者纳入研究,术前3h经纤维肠镜、肛镜于病灶周围粘膜下注入99mTc标记的右旋糖苷,术中在病灶周围浆膜下注入亚甲蓝,探测仪检测放射性高出背景组织10倍以上或(和)蓝染的淋巴结视为结直肠癌的前哨淋巴结(sentinel lymphnode,SLN),行常规病理检查,分别计算前哨淋巴结诊断结直肠癌区域淋巴结转移状态、假阴性率等,并根据SLN活检结果决定结直肠癌的手术方式。结果:本组结直肠癌SLN的检出成功率为80%(16/20),每例检出1～3个,平均2.4个/例,SLN的转移率为37.5%(18/48);诊断敏感性80%(16/20);诊断准确率83.3%(15/18);假阴性率20%(4/20)。结论:前哨淋巴结活检术适合于结直肠癌,联合示踪法检测结直肠癌前哨淋巴结可判断区域淋巴结的转移状态,并可用于指导结直肠癌淋巴结清扫范围。     

前哨淋巴结活检术在结直肠癌中的应用研究

目的：探讨结直肠癌中前哨淋巴结定位活检术的适用性。方法：20例结直肠癌患者纳入研究,术前3h经纤维肠镜、肛镜于病灶周围粘膜下注入99mTc标记的右旋糖苷,术中在病灶周围浆膜下注入亚甲蓝,探测仪检测放射性高出背景组织10倍以上或（和）蓝染的淋巴结视为结直肠癌的前哨淋巴结（sentinel lymphnode,SLN）,行常规病理检查,分别计算前哨淋巴结诊断结直肠癌区域淋巴结转移状态、假阴性率等,并根据SLN活检结果决定结直肠癌的手术方式。结果：本组结直肠癌SLN的检出成功率为80%（16/20）,每例检出1～3个,平均2.4个/例,SLN的转移率为37.5%（18/48）;诊断敏感性80%（16/20）;诊断准确率83.3%（15/18）;假阴性率20%（4/20）。结论：前哨淋巴结活检术适合于结直肠癌,联合示踪法检测结直肠癌前哨淋巴结可判断区域淋巴结的转移状态,并可用于指导结直肠癌淋巴结清扫范围。     

逆转录聚合酶链反应检测结直肠癌淋巴结微转移的临床意义

背景与目的:结直肠癌淋巴结微转移灶是否有预测预后价值目前尚有争议,本文对结直肠癌患者淋巴结微转移情况进行逆转录聚合酶链反应(RT-PCR)检测,研究微转移对临床分期和预后的影响。方法:用RT-PCR技术检测56例结直肠癌患者肠旁及系膜淋巴结中细胞角质蛋白CK20mRNA,揭示微转移灶的存在,并与常规病理苏木精伊红(HE)染色和免疫组化染色结果进行比较;分析HE染色和RT-PCR检测结果对判定临床病理分期和统计生存率的影响。结果:共检测432个淋巴结,HE染色、免疫组化染色和RT-PCR法淋巴结转移检出率分别为57.2%、62.3%和73.1%。HE染色和免疫组化的检出率无显著性差异(P>0.05),而HE染色和RT-PCR法检测结果有显著性差异(P<0.05)。56例患者中,按HE染色结果确定的PN0、PN1和PN2期,其5年无复发转移生存率分别为80%、60%和50%;通过RT-PCR技术检测,升级后的PN0、PN1和PN2期5年无复发转移生存率分别为100%、61.9%和55.6%,两种方法分析的结果有显著性差异(P<0.05)。结论:HE染色未确切指出淋巴结微转移癌;RT-PCR法检测CK20mRNA可以推断淋巴结微转移癌的存在,从而有助于确定结直肠癌临床分期和预测预后。     

IHC和PCR法联合检测乳腺癌前哨淋巴结的微转移

目的:探讨采用免疫组织化学(IHC)和RT-PCR方法同时检测乳腺癌前哨淋巴结(SLN)微转移的灵敏度及其临床价值。方法:全身麻醉后用1%异硫蓝行腋窝前哨淋巴结活检(SLNB),即刻取1/2SLN投入液氮中保存,然后行乳腺癌改良根治术。经常规病理切片证实包括SLN在内的腋窝淋巴结阴性患者62例,同时采用IHC和RT-PCR方法检测SLN微转移。结果:62例患者中,13例(20·97%)SLN有KT19mRNA表达,其中7例(11·29%)IHC法也检出KT19阳性细胞,表明RT-PCR法灵敏度较IHC法更高,两者差异有统计学意义,χ2=4·1667,P=0·0412;两者检测结果有较好的一致性,kappa=0·6483。另外,比较临床生物学特性和SLN微转移之间关系,未发现有相关因素。结论:常规检查未发现远处和腋窝淋巴结转移的乳腺癌患者,经IHC和RT-PCR法对SLN有针对性的检测可发现微转移的存在,其中RT-PCR方法较IHC更灵敏。     

淋巴结微转移检测对结直肠癌病理分期的影响

目的:探讨淋巴结微转移对结直肠癌患者病理分期的影响。方法:对2003年3月至2008年10月间手术切除的60例结直肠癌患者的990枚淋巴结进行常规HE染色和CK20免疫组化检查,检测结直肠癌的淋巴结转移,并对结果进行统计学分析。结果:应用HE染色法淋巴结转移检出率27.2%(269/990),CK20免疫组化淋巴转移检测率为37.2%(368/990),两者差异有统计学意义(P<0.05)。99枚淋巴结检出有微转移,11例TNM分期提高,HE染色重新分期率为18.3%(11/60)。结论:CK20免疫组化的检测可以显著提高淋巴结转移的检出率,有助于更准确地进行结直肠癌的临床病理分期。     

应用RT-PCR检测CK19表达提高乳腺癌前哨淋巴结微转移检出的研究

目的:评价乳腺癌前哨淋巴结活检的临床意义,应用细胞角蛋白cytokeratin19(CK19)RT-PCR检测法提高其的敏感性及准确率.方法:随机选取1999年10月至2003年3月住院的乳腺癌患者73例,其中2001年1月至2003年3月的29例,对前哨淋巴结(SLN)进行了CK19 RT-PCR检测.采用专利兰进行SLN识别.RT-PCR法检测CK19 mRNA表达.结果:SLN识别成功67例,成功率为91.8%(67/73).对上述67例的SLN转移的常规病理诊断的敏感度68.8%(22/32),准确率85.1%(57/67).在行CK19 RT-PCR的29例中,检测出常规病理未检出的微小转移病例8例.敏感度90.0%(18/20),准确率93.1%(27/29).结论:前哨淋巴结活检可有效判断乳腺癌腋淋巴结转移状态,应用CK19 RT-PCR检测SLN的微转移,可提高敏感度及准确率.     

Significance of Sentinel Lymph Node Biopsy and Immunohistochemistry in Diagnosis and Staging and staging of Stage-cNO Oral Squamous Cell Carcinoma

OBJECTlVE To assess the significance of sentinel lymph node biopsy (SLNB),serial section and cytokeratin immunohistochemical staining in the diagnosis and staging of Stage-cNO oral squamous cell carcinoma (OSCC).METHODS A blue stain,99mTc.dextran SPECT lymphoscintigrapgy and intraoperative γ-ray probes were used to examine the sentinel nodes in 31 cases with Stage-cNO oral cancer.The H&E staining and a cytokeratin AE1/AE3 immunohistochemistry (IHC) assessment,with serial sections,were conducted to provide results obtained from a routine pathological examination of lymph nodes.The value of the routine pathological examination of the sentinel lymph node (SLN),serial sections and IHC determination for cervical Iymph node metastasis of Stage-cNO OSCC was appraised.RESULTS A total of 45,55 and 51 SLNs were examined in 25 (80%),31 (100%) and 30(96.5%) of the cases,by using the blue stain,γ-ray probes,and SPECT lymphoscintigraphy,respectively.The average SLNs found in each case of the groups was 1.4(1 to 3)and there were 1,302 non-NSLNs.Six positive SLN metastases were detected by routine pathological examination,among which 1 case was found to be an accompanied positive metastasis of non-SLN.One positive SLN metastasis was found after examination of serial sections plus routine H&E staining and 2 were detected using serial sections plus AE3 immunohistochemical staining methods.No positive NSLNs were found in the study.CONCLUSION In order to make more progress in accurate SLNB diagnosis,serial sections and OHC (AE1/AE3) methods can be used for examination of the micrometastases which are difficult to identify by routine pathological sections and H&E staining.node,micrometastasis,serial sections,immunohistochemistry.     

One-step nucleic acid amplification for intraoperative detection of lymph node metastasis in breast cancer patients.

PURPOSE: Detection of sentinel lymph node (SLN) metastasis in breast cancer patients has conventionally been determined by intraoperative histopathologic examination of frozen sections followed by definitive postoperative examination of permanent sections. The purpose of this study is to develop a more efficient method for intraoperative detection of lymph node metastasis. EXPERIMENTAL DESIGN: Cutoff values to distinguish macrometastasis, micrometastasis, and nonmetastasis were determined by measuring cytokeratin 19 (CK19) mRNA in histopathologically positive and negative lymph nodes using one-step nucleic acid amplification (OSNA). In an intraoperative clinical study involving six facilities, 325 lymph nodes (101 patients), including 81 SLNs, were divided into four blocks. Alternate blocks were used for the OSNA assay with CK19 mRNA, and the remaining blocks were used for H&E and CK19 immunohistochemistry-based three-level histopathologic examination. The results from the two methods were then compared. RESULTS: We established CK19 mRNA cutoff values of 2.5 x 10(2) and 5 x 10(3) copies/muL. In the clinical study, an overall concordance rate between the OSNA assay and the three-level histopathology was 98.2%. Similar results were obtained with 81 SLNs. The OSNA assay discriminated macrometastasis from micrometastasis. No false positive was observed in the OSNA assay of 144 histopathologically negative lymph nodes from pN0 patients, indicating an extremely low false positive for the OSNA assay. CONCLUSION: The OSNA assay of half of a lymph node provided results similar to those of three-level histopathology. Clinical results indicate that the OSNA assay provides a useful intraoperative detection method of lymph node metastasis in breast cancer patients.     

早期宫颈癌前哨淋巴结中高危型HPV16/18 DNA表达的检测及临床意义

目的：探讨早期宫颈癌患者前哨淋巴结（SLN）中人乳头状瘤病毒（HPV）16/18 DNA表达检测对于微转移的临床意义。方法选取72例早期宫颈癌患者,予患者均行广泛性子宫切除加双侧盆腔淋巴结清扫术,术中采用染料法识别SLN的宫颈癌患者有46例,应用基因检测法（FQ-PCR）检测SLN中HPV16/18 DNA阳性表达情况,并分析其与各种临床病理因素的关系;对SLN病理阴性的33例患者进行长期随访,分析SLN中HPV16/18 DNA阳性与淋巴结转移的关系。结果46例宫颈癌患者SLN中HPV16/18 DNA阳性表达者共22例,其中13例淋巴结病理阳性患者中有10例阳性,而33例淋巴结病理阴性患者中仅12例阳性（P=0.013）;46例患者共检出前哨淋巴结102枚,均用FQ-PCR法检测HPV16/18 DNA,结果13例淋巴结病理阳性患者检出的37枚SLN中有29枚HPV16/18 DNA阳性,而33例淋巴结病理阴性患者检出的65枚SLN中仅有36枚阳性（P=0.033）;分析46例成功检出SLN的早期宫颈癌患者的临床资料,发现SLN中HPV16/18 DNA阳性表达仅与临床分期有关,具有统计学意义（P=0.034）;长期随访33例SLN病理阴性的患者,发现HPV16/18 DNA阳性的患者复发率高于HPV16/18 DNA阴性的患者,具有统计学意义（P=0.02）。结论检测宫颈癌SLN组织中HPV16/18 DNA表达可能是预测早期宫颈癌淋巴结微转移的可行方法。     

早期非小细胞肺癌前哨淋巴结微转移检测的应用研究

目的 目前,尚无检测技术可准确判断肺癌前哨淋巴结(sentinel lymph node,SLN)微转移.本研究探讨CK19和MAGE A3表达与非小细胞肺癌(non-small cell lung cancer,NSCLC) SLN微转移的相关性及临床价值.方法 选择山东大学附属山东省肿瘤医院胸外科32例接受手术治疗的临床Ⅰ～ⅡA期NSCLC患者,术中联合应用染色法(异舒泛蓝溶液)和放射同位素法(99 Tc硫胶体检测)找寻SLN,并采用免疫组化技术检测SLN及非前哨淋巴结(non-sentinel lymph node,nowSLN)中CK19和MAGE-A3抗体的表达.结果 32例患者均检测出SLN,共清除淋巴结598枚,其中SLN 103枚,non-SLN 495枚.平均每例患者清除淋巴结(18.69±8.13)枚,清除SLN(3.22±1.74)枚.免疫组化法检测到20例患者44枚SLN中CK19表达阳性,19例患者31枚SLN中MAGE-A3抗体表达阳性.SLN免疫组化检查阳性率为42.72％,明显高于常规HE染色的阳性率(25.24％),P=0.01.SLN的阳性表达率与临床病理分期有关,P＜0.05;而与性别、年龄、肿瘤部位、分化程度、肿瘤大小和肿瘤类型无关,P＞0.05.结论 CK19和MAGMA3是判断淋巴结微转移较好的分子标志物,通过免疫组化技术检测SLN中CK19和MAGE-A3表达有助于评估区域淋巴结微转移状况.     

乳腺癌前哨淋巴结活检40例报告

目的 :通过对乳腺癌前哨淋巴结 (SLN)活检 ,探讨其对腋淋巴结转移情况预测的准确性。方法 :采用亚甲蓝染料法对 4 0例乳腺癌行腋窝蓝染淋巴结活检 ,后行常规腋窝淋巴结清除 (ALND) ,两标本均送病理检查。结果 :全组 4 0例患者检出SLN 38例 ,2例未找到SLN ,检出率为 95 % ( 38/ 4 0 ) ,有 8例SLN为阳性 ,1例SLN为假阴性 ,腋窝淋巴结 (ALN)有 9例转移 ,SLN与ALN病理检查完全符合者 37例 ,准确率为 92 5 % ( 37/ 38) ;灵敏度为 88 9% ( 8/ 9) ;假阴性率为 11 1% ( 1/ 9)。结论 :亚甲蓝染色法能准确地鉴别SLN及预测乳腺癌腋窝淋巴结状态     

Significance of Sentinel Lymph Node Biopsy and Immunohistochemistry in Diagnosis and Staging of Stage-cNO Oral Squamous Cell Carcinoma

OBJECTIVE To assess the significance of sentinel lymph node biopsy （SLNB）, serial section and cytokeratin immunohistochemical staining in the diagnosis and staging of Stage-cNO oral squamous cell carcinoma （OSCC）, METHODS A blue stain, 99mTc-dextran SPECT lymphoscintigrapgy and intraoperative y-ray probes were used to examine the sentinel nodes in 31 cases with Stage-oNO oral cancer, The H＆E staining and a cytokeratin AE1/ AE3 immunohistochemistry （IHC） assessment, with serial sections, were conducted to provide results obtained from a routine pathological examination of lymph nodes, The value of the routine pathological examination of the sentinel lymph node （SLN）, serial sections and IHC determination for cervical lymph node metastasis of Stage-cN0 OSCC was appraised, RESULTS A total of 45, 55 and 51 SLNs were examined in 25 （80%）, 31 （100%） and 30 （96,5%） of the cases, by using the blue stain, y-ray probes, and SPECT lymphoscintigraphy, respectively, The average SLNs found in each case of the groups was 1,4 （1 to 3） and there were 1,302 non-NSLNs, Six positive SLN metastases were detected by routine pathological examination, among which 1 case was found to be an accompanied positive metastasis of non-SLN, One positive SLN metastasis was found after examination of serial sections plus routine H＆E staining and 2 were detected using serial sections plus AE3 immunohistochemical staining methods, No positive NSLNs were found in the study, CONCLUSION In order to make more progress in accurate SLNB diagnosis, serial sections and IHC （AE1/AE3） methods can be used for examination of the micrometastases which are difficult to identify by routine pathological sections and H＆E staining.     

Intraoperative, radio-guided sentinel lymph node mapping in laparoscopic lymph node dissection for Stage I testicular carcinoma

BACKGROUND: The management of regional lymph nodes in patients with clinical Stage I testicular carcinoma is a controversial problem. The authors investigated the feasibility and accuracy of radio-guided mapping of sentinel lymph nodes (SLNs) for men with clinical Stage I testicular tumors. METHODS: Twenty-two patients with clinical Stage I testicular carcinoma were enrolled in the study. One day before surgery, (99m)Technetium-labeled phytate was injected around the testicular tumor. After undergoing radical orchiectomy, patients underwent laparoscopic retroperitoneal lymph node dissection (L-RPLND). All radioactive lymph nodes were marked in the L-RPLND procedure, and three-dimensional SLN maps were made. All resected lymph nodes were evaluated by routine histopathologic examination, and the clinical significance of intraoperative SLN mapping was evaluated. RESULTS: SLNs were detected in 21 of 22 patients (95%). Nearly all SLNs were detected at the ventral or lateral side of the vena cava or at the aorta between the levels of the aortic bifurcation. All SLNs were detected easily in a surgical procedure. Only 1 radio-positive area per patient was identified in 15 patients, and approximately 2-4 positive areas were detected in 6 patients. Two patients had micrometastasis only in SLNs. In 2 patients who had seminoma, lymph node recurrences (at the level of the renal vein and in the obturator lymph node area) occurred at 10 months and 20 months after surgery. CONCLUSIONS: Radio-guided mapping of SLNs with laparoscopy was feasible, and nearly all SLNs were detected accurately by the procedure. In the near future, the standard retroperitoneal lymph node dissection may be avoided in most patients with clinical Stage I testicular carcinoma by utilizing focused examination of SLNs.