Flow cytometric analysis of DNA content in colorectal polyps resected at endoscopy

Samples from 64 consecutive resected colorectal polyps were preserved in liquid nitrogen and analyzed by flow cytometry to assess the nuclear DNA content, which was compared to the histopathologic findings. The frequency of aneuploidy in these nonselected polyps was 17.2%. There was a significant correlation between the diameter of the polyp and the frequency of aneuploidy (P = .04), with all aneuploid polyps being greater than or equal to 10 mm. Similarly, aneuploidy was significantly more frequent for polyps that were both greater than or equal 20 mm and showed at least severe dysplasia (P = .02). On the other hand, there was no correlation between the ploidy and the gross histopathologic type (tubular, tubulovillous or villous), and the proliferation index did not correlate with any of the parameters studied.     

Ratio of Metastatic to Examined Lymph Nodes,a Helpful Staging System and Independent Prognostic Factor of Esophagogastric Junction Cancer

Background

The incidence of the esophagogastric junction cancer is growing rapidly. The purpose of this study is to clarify the outcome of the ratio between metastatic and examined lymph nodes in esophagogastric junction cancer patients with or without 7 examined lymph nodes.

Methods

A total of 3,481 patients who underwent operation are identified from the Surveillance, Epidemiology, and End Results database. Different lymph nodes resected groups are analyzed to test the lymph nodes ratio factor.

Results

There are 2522 patients with 7 or more lymph nodes resected and 959 patients with less than 7 lymph nodes resected. Lymph nodes ratio and lymph node involvement are independent prognostic factors. But the lymph nodes ratio categories have a better prognostic value than the lymph node involvement categories. Compared with lymph node involvement categories, lymph nodes ratio categories represent patients with more homogeneous overall survival rate.

Conclusions

This study defines that the lymph nodes ratio is an independent prognostic factor for esophagogastric junction cancer. The lymph nodes ratio can prevent stage migration and may be a helpful system to predict the prognosis of esophagogastric junction cancer patients.     

结直肠癌mP53 和PCNA 表达的相关性及临床意义

目的:探讨结直肠癌中突变型P53基因(mP53)和增殖细胞核抗原(PCNA)表达的相关性及临床意义。方法:应用免疫组化二步法,检测60例结直肠癌组织及20例正常肠粘膜中mP53、PCNA的表达,结合临床病理资料进行统计分析。结果:60例结直肠癌中mP53阳性表达率65.0%,PCNA阳性表达率78.3%,20例正常肠粘膜中mP53、PCNA表达均为阴性(P<0.05)。mP53和PCNA阳性表达率在低分化组、浆膜层浸润组、淋巴结转移组均较高(P均<0.05)。mP53和PCNA表达呈正相关(r=0.58,P<0.05)。结论:mP53和PCNA在结直肠癌中表达均增高,二者与结直肠癌病理学分级、浸润深度和淋巴结转移有关,可作为判断结直肠癌预后的参考指标。     

Gradient of nuclear volume changes in breast cancer and its relation to prognosis

OBJECTIVE: In a study of 112 cases of invasive breast cancer, the prognostic value of the gradient of nuclear volume (NV) changes was examined. STUDY DESIGN: On conventional histologic slides of surgically resected specimens, we studied the nuclear changes from early to advanced. The former was represented by the noninvasive area in the tumor periphery and the latter by the invasive area with true neoplastic stroma. We analyzed the difference in NV between the noninvasive area in the tumor periphery (NVcis) and the invasive area where the nuclei are large (NVlarge), a zone we assessed to have the greatest cancer progression. Based on these measurements, we defined NV gradient index (NVG index) as the ratio of NVlarge to NVcis. RESULTS: The NVG index in cases of cancer death (4.33 +/- 1.70, n = 35) was significantly larger than in relapsing survivors (2.76 +/- 1.31, n = 13, P = .002) and in relapse-free survivors (2.17 +/- 1.60, n = 64, P < .0003). In the group of cancer deaths, the correlation coefficient between NVG index and survival was -0.398 (0 < P < .02). Even in cases of stage I, NVG index in the group of cancer deaths (5.68 +/- 1.4) was significantly larger than in relapsing survivors (2.42 +/- 1.2) or relapse-free survivors (1.74 +/- 0.55). By multivariate analysis, NVG index was independently prognostic for disease-specific survival (P = .0001). CONCLUSION: The NVG index contributes to discrimination between possible survivors and cancer deaths. Though there are exceptions, cases with a small NVG index can be expected to survive for a longer period even after a relapse.     

Increased Lymph Node Yield in Colorectal Cancer Is Not Necessarily Associated with a Greater Number of Lymph Node Positive Cancers

The presence of lymph node metastasis is a key prognostic factor in colorectal cancer and lymph node yield is an important parameter in assessing the quality of histopathology reporting of colorectal cancer excision specimens. This study assesses the trend in lymph node evaluation over time in a single institution and the relationship with the identification of lymph node positive tumours. It compares the lymph node yield of a contemporary dataset compiled from the histopathology reports of 2178 patients who underwent surgery for primary colorectal cancer between 2005 and 2012 with that of a historic dataset compiled from the histopathology reports of 1038 patients who underwent surgery for colorectal cancer at 5 yearly intervals from 1975 to 2000. The mean lymph node yield was 14.91 in 2005 rising to 21.38 in 2012. In 2012 92.9% of all cases had at least 12 lymph nodes examined. Comparison of the mean lymph node yield and proportion of Dukes C cases shows a significant increase (Pearson correlation = 0.927, p = 0.001) in lymph node yield while there is no corresponding significant trend in the proportion of Dukes C cases (Pearson correlation = −0.138, p = 0.745). This study shows that there is increasing yield of lymph nodes from colorectal cancer excision specimens. However, this is not necessarily associated with an increase number of lymph node positive cancers. Further risk stratifying of colorectal cancer requires consideration of other pathological parameters especially the presence of extramural venous invasion and relevant biomarkers.     

Clinical Features in Differentiated Thyroid Carcinoma Stratified By Lymph Node Status

Objective: Cervical lymph node (CLN) metastases (mets) often occur in differentiated thyroid cancer (DTC), especially in the central compartment, and are a major predictor of local recurrence. We examined clinical endpoints in three groups of patients based on status of lymph node involvement: those with definite lymph node involvement (N1), negative lymph nodes (N0), and no lymph nodes resected (Nx). We correlated these endpoints with clinical and pathologic features of these patients.Methods: Medical records of 261 patients with DTC who underwent thyroidectomy between 2006 and 2018 at our center were reviewed. Lymph node status of patients was categorized based on American Joint Committee on Cancer (AJCC) 8th edition criteria as N1, N0, and Nx. We performed statistical analysis to assess the differences among these groups, using one-way analysis of variance. When significant differences were found, pairwise comparisons were conducted among the three groups. Statistical significance was defined as 2-tailed P<.05 for all tests.Results: There were significant differences among the groups in tumor multicentricity, tumor category/size, AJCC stage, and the presence of thyroglobulin auto-antibodies (TgAbs). There were no difference in age, gender, or histopathology. N1 patients had a higher incidence of multicentricity, larger tumor sizes, and were more likely to have elevated TgAbs. There were no significant differences between the N0 and Nx groups.Conclusion: This study shows that larger and multi-centric tumors are associated with increased likelihood of CLN mets in DTC. We suggest increased vigilance for CLN mets in tumors >2 cm, multicentric tumors, and patients with elevated TgAbs.Abbreviations: AJCC = American Joint Committee on Cancer; CLN = cervical lymph node; DTC = differentiated thyroid cancer; FTC = follicular thyroid cancer; mets = metastases; N0 = no cancer in any lymph nodes; N1 = cancer in a lymph node; N1a = cancer in a central compartment lymph node; N1b = cancer in a lateral neck lymph node; Nx = lymph nodes not resected or examined; PTC = papillary thyroid cancer; TgAb = thyroglobulin auto-antibody     

The number distribution for involved lymph nodes in cancer

The number of involved lymph nodes exhibits considerable heterogeneity within populations. Here, the implications of population heterogeneity are explored with respect to the kinematics of nodal metastases. Data from the National Cancer Institute's Surveillance, Epidemiology, and End Results program for 224656 breast, 12404 gastric, 18015 rectal, 4117 cervical and 2443 laryngeal cancers as well as 9118 melanomas were used to construct frequency distributions for the number of involved nodes which were then fitted to the negative binomial distribution. The negative binomial distribution described the heterogeneity in nodal involvement well. The patterns of nodal involvement can be explained by either of two models: one where involved nodes could seed further nodal metastases, the other where the number of nodal metastases in any individual was randomly distributed, with the deviations between patients accounted for by population heterogeneity. Since the number of sampled nodes similarly approximated a negative binomial distribution, random involvement with superimposed population heterogeneity would more credibly explain both sets of observations.     

Cytophotometric and electron microscopic studies of chronic human skin lymphomas. II. Research on lymph node involvement

Results of electron microscopic and cytophotometric studies of biopsy material from lymph nodes of patients with cutaneous lymphomas with low degree of malignancy are discussed, with special reference to early diagnostic criteria of the disease. Specific characteristics of tumor tissue involve the presence of atypical lymphocytes with marginal condensed chromatin extrusions of nuclear material, deep invaginations of the nuclear membrane, nuclear pockets, excess of the mean DNA content per nucleus above the diploid standard level, more than 5% of nuclei being in the hyperdiploid area. Electron microscopic and cytophotometric methods allowed to diagnose the tumor injury of lymph nodes, when the traditional histological techniques revealed no signs of malignancy.     

Correlation of N-myc downstream-regulated gene 1 subcellular localization and lymph node metastases of colorectal neoplasms

In colorectal neoplasms, N-myc downstream-regulated gene 1 (NDRG1) is a primarily cytoplasmic protein, but it is also expressed on the cell membrane and in the nucleus. NDRG1 is involved in various stages of tumor development in colorectal cancer, and it is possible that the different subcellular localizations may determine the function of NDRG1 protein. Here, we attempt to clarify the characteristics of NDRG1 protein subcellular localization during the progression of colorectal cancer. We examined NDRG1 expression in 49 colorectal cancer patients in cancerous, non-cancerous, and corresponding lymph node tissues. Cytoplasmic and membrane NDRG1 expression was higher in the lymph nodes with metastases than in those without metastases (P < 0.01). Nuclear NDRG1 expression in colorectal neoplasms was significantly higher than in the normal colorectal mucosa, and yet the normal colorectal mucosa showed no nuclear expression. Furthermore, our results showed higher cytoplasmic NDRG1 expression was better for differentiation, and higher membrane NDRG1 expression resulted in a greater possibility of lymph node metastasis. These data indicate that a certain relationship between the cytoplasmic and membrane expression of NDRG1 in lymph nodes exists with lymph node metastasis. NDRG1 expression may translocate from the membrane of the colorectal cancer cells to the nucleus, where it is involved in lymph node metastasis. Combination analysis of NDRG1 subcellular expression and clinical variables will help predict the incidence of lymph node metastasis.     

结直肠癌淋巴结微转移灶的临床病理研究

摘要目的：探讨结直肠癌淋巴结微转移灶的特点及其临床病理意义。方法：对我院2001年1月-2006年10月收治的56例无组织学淋巴结转移的结直肠癌患者的650个淋巴结进行微转移灶的免疫组织化学和组织学检测,并分析微转移灶与患者临床病理特征之间的关系。结果-通过对41,zm厚淋巴结连续切片进行观察,连续切片五片的微转移灶检出率较一片和两片的的微转移灶检出率显著升高（P〈0．05）。按解剖部位分组的患者微转移灶阳性率1组有66．67％（16／24）,2组有60％（9／15）,3组有41．2％（7／17）,无显著性差别（P〉0．05）。微转移灶与无组织学淋巴结转移的结直肠癌患者的肿瘤浸润深度（P=0．013）和肿块大小（P=0．037）存在显著性相关,但有微转移灶和无微转移灶的无组织学淋巴结转移的结直肠癌患者总体生存状况比较无显著差异（P〉0．05）。结论：微转移灶在无组织学淋巴结转移的结直肠癌淋巴结中较常见,与肿瘤大小和浸润深度有关,但与患者的预后无显著相关性。     

Adjuvant Treatment of Colorectal Cancer: A Review

Surgical operation remains the most effective method of treatment for patients with cancer of the large bowel. However, innovative surgical techniques have not improved survival rates for colorectal cancer in 25 years.Attempts at increasing survival with chemotherapy as an adjunct to surgical procedures remain inconclusive and controversial. Many adjuvant chemotherapy trials have failed to recognize those prognostic factors—such as nodal involvement, serosal penetration, vascular or perineural invasion, and microscopic invasion at margins of resection—that characterize certain patients at high risk for recurrent cancer. Failure to include only high risk patients in adjuvant chemotherapy is, in part, responsible for the lackluster performance to date.For rectal cancer, preoperative irradiation increases the chances of cure with surgical operation by reduction of pathologic staging, but it has not increased survival in patients with persistent nodal involvement.Immunotherapy is a possibly valuable method of treatment; however, it is clinically untested. An adjuvant immunotherapy protocol for high risk patients is described.     

64排多层螺旋CT对直肠癌淋巴结转移的诊断研究

目的:通过对直肠癌患者原发肿瘤周围脂肪间隙内淋巴结的影像学分析,来评价64排增强螺旋CT对直肠癌淋巴结转移的诊断价值。方法:收集我院经术后病理证实的102例直肠癌患者的临床及影像学资料进行回顾性分析,通过多平面成像,记录直肠周围脂肪间隙内淋巴结的CT表现,包括淋巴结大小、边缘及强化程度。以病理结果为准确定转移性和非转移性淋巴结,采用双盲法。结果:单因素分析结果显示淋巴结大小、边缘及强化方式与阳性率有显著的相关性(P0.05),多因素分析结果显示各相关因素有显著诊断意义的是淋巴结大小、边缘及强化方式,其敏感性分别为94.7%、87.6%、56.9%,特异性分别为69.3%、51.6%、62.3%。结论:综合分析原发肿瘤周围脂肪间隙内淋巴结的表观情况,有助于提高对淋巴结转移的判断。     

Correlation of anemia with infradiaphragmatic involvement in Hodgkin's disease and other malignant lymphomas

A staging laparotomy and splenectomy were performed in 41 patients with Hodgkin''s disease and 11 with other malignant lymphomas. There was a significant correlation (P = 0.025) between the presence of unexplained anemia and involvement of the spleen or abdominal lymph nodes by tumour. The anemias were of mild degree; hemolysis was documented in three and iron deficiency in four, while 21 cases were unexplained. Bone marrow was not involved by lymphoma in this series. The complication rate in exploratory laparotomy was higher than previously reported. Severe complications were observed in 17% of these patients while another 15% had minor complications. The association we have discovered may be helpful in the staging of patients who cannot tolerate an operative procedure. The absence of infradiaphragmatic involvement is suggested in the presence of normal hemoglobin concentrations.     

Cell kinetics analysis in stage I endometrial carcinoma as determined by image cytometry and immunohistochemistry. Correlation with clinicopathologic characteristics

OBJECTIVE: To test the significance of various proliferative indices in endometrial carcinoma (EC) since previous investigations failed to yield consistent results that would establish them as factors of clinical importance. STUDY DESIGN: Seventy patients with EC were evaluated for various proliferative indices (mitotic index, Ki-67 index, argyrophilic nucleolar organizer (AgNOR) number and area per nucleus, and p53 protein expression) in relation to image cytometry (nuclear area, diameter and roundness) and standard clinicopathologic features (age, histologic type and grade, and depth of invasion). We also tested the proliferation index (PI), which combines the Ki-67 index and AgNOR area in Ki-67-positive nuclei. Slides from each case were double stained for Ki-67 antigen and AgNOR proteins for this purpose. RESULTS: Mitotic counts were significantly higher in papillary-serous (vs. endometrioid) tumors (P = .0001), high grade (vs. low grade) tumors (P = .0001), deeply invasive (P = .017) and p53-positive tumors (P = .017). AgNOR counts correlated only with age (higher in older women, P = .002), while the PI correlated with mitotic counts (P = 0.28) and marginally with depth of invasion (P = .06). Morphometric variables were associated just with histologic type and grade. p53 Protein was expressed exclusively in invasive tumors and was related strongly to histologic type (P = .0029) and grade (P = .0001). CONCLUSION: Our data reestablish the value of classic histopathologic features (mitotic index, histologic type and grade) as the most important tools for EC evaluation. In addition, we suggest that p53 immunostaining may be used for predicting aggressive behavior in EC.     

Nuclear planimetry in renal-cell tumors

A morphometric study using conventional planimetry was carried out in 73 renal-cell tumors (64 adenocarcinomas and 9 adenomas) in order to correlate the objective nuclear measurements with some histologic and clinical data believed to have prognostic value. The use of a discriminant function to correctly separate adenomas and low-grade adenocarcinomas was also investigated. There was a strong association between the nuclear grade and the following parameters: nuclear area, nuclear major diameter and nuclear elongation. There was also a relationship between an adverse outcome and the major nuclear diameter and nuclear elongation. In a univariate survival analysis, only the clinicopathologic stage, the nuclear grade and the histologic presence of a pseudosarcomatous cell population had predictive value. Multivariate survival analysis using Cox models showed that the clinicopathologic stage, major nuclear diameter, presence of anaplastic cells and nuclear area had predictive value. Although the minor nuclear diameter reached a significant level, a discriminant function that would correctly separate adenomas and low-grade renal-cell carcinomas could not be formulated.     

A basal membrane-like structure surrounding tumour nodules may prevent intraepithelial leucocyte infiltration in colorectal cancer

Epithelial tumours consist of an epithelial compartment and a stromal compartment, which are sometimes separated by a basal membrane-like structure. We sought to determine whether these factors have prognostic value in 84 curatively resected stage II and III colorectal cancer by immunohistochemically staining tumours for leucocytes (CD45) and extracellular matrix, and to assess the presence of a basal membrane-like structure. Leucocyte infiltration was also assessed in hematoxylin-eosin (HE) stained sections. Most leucocytes were located in the tumour stroma. A relatively high intraepithelial leucocyte infiltration was significantly correlated with a lower level of tumour recurrence (P=0.03) and a longer disease-free survival (P=0.05), whereas leucocytes located in the tumour stroma (P=0.92) or at the advancing margin (p=0.06) were not. Intraepithelial leucocyte infiltration was also significantly correlated with leucocyte infiltration in the tumour stroma (P=0.02) and at the advancing tumour margin (P=0.005), and as assessed in HE-stained tumour sections (P=0.05), but each of these parameters on its own did not have a prognostic value in predicting disease-free survival. Moreover, the presence of a basal membrane-like structure surrounding the tumour epithelium was inversely correlated with the number of intraepithelial leucocytes (P=0.05), suggesting that this membrane-like structure functions as a barrier to intraepithelial leucocyte infiltration. We conclude that leucocytes must be in the direct vicinity of tumour cells to affect tumour growth. The presence of an extracellular matrix barrier seems to prevent this interaction.     

Computerized morphometric image analysis of cytologic nuclear parameters in breast cancer

The value of nuclear morphometry in the preoperative fine needle aspiration (FNA) cytologic diagnosis of mammary lesions was investigated and correlated with the lymph node status of the patients. The subjects consisted of four groups of patients: 49 with invasive ductal carcinomas (18 with no positive nodes, 16 with one to three positive nodes and 15 with four or more positive nodes) and 14 patients with benign lesions. The FNA specimens were smeared onto slides and stained by the May-Grünwald-Giemsa technique. The area, perimeter and maximum diameter of 100 randomly chosen nuclei were both measured with the IBAS image analysis system and semiquantitatively estimated with an eyepiece micrometer. For all three parameters, significant differences were found between benign and malignant lesions. The mean nuclear perimeter allowed the morphometric discrimination between all four groups with statistical significance; nuclear area and maximum diameter did not discriminate patients with invasive carcinoma and one to three positive nodes from those with no positive nodes or more than three positive nodes. Morphometry proved to be far superior to eyepiece measurements with respect to accuracy and reproducibility of the results. The results suggest that nuclear perimeter can be used as an additional parameter not only for the FNA cytologic diagnosis of breast cancer, but also for the estimation of patients' prognosis.     

Sentinel lymph node biopsy for melanoma: experience with 234 consecutive procedures

Sentinel lymph node biopsy is increasingly used to identify occult metastases in regional lymph nodes of patients with melanoma. Selection of patients for sentinel lymph node biopsy and subsequent lymphadenectomy is an area of debate. The purpose of this study was to describe a large clinical series of these biopsies for cutaneous melanoma and to identify patients most likely to gain useful clinical information from sentinel lymph node biopsy. The Indiana University Melanoma Program computerized database was queried to identify all patients who underwent this procedure for clinically localized cutaneous melanoma. It was performed using preoperative technetium Tc 99m lymphoscintigraphy and isosulfan blue dye. Pertinent demographic, surgical, and histopathologic data were recorded. Univariate and multivariate logistic regression and classification table analyses were performed to identify clinical variables associated with sentinel node and nonsentinel node positivity. In total, 234 biopsy procedures were performed to stage 291 nonpalpable regional lymph node basins. Mean Breslow's thickness was 2.30 mm (2.08 mm for negative sentinel lymph node biopsy, 3.18 mm for positive). The mean number of sentinel nodes removed was 2.17 nodes per basin (range, 1 to 8). Forty-seven of 234 melanomas (20.1 percent) and 50 of 291 basins (17.2 percent) had a positive biopsy. Positivity correlated with AJCC tumor stage: T1, 3.6 percent; T2, 8.1 percent; T3, 27.4 percent; T4, 44 percent. By univariate logistic regression, Breslow's thickness (p = 0.003, continuous variable), ulceration (p = 0.003), mitotic index > or = 6 mitoses per high power field (p = 0.008), and Clark's level (p = 0.04) were significantly associated with sentinel lymph node biopsy result. By multivariate analysis, only Breslow's thickness (p = 0.02), tumor ulceration (p = 0.02), and mitotic index (p = 0.02) were significant predictors of biopsy positivity. Classification table analysis showed the Breslow cutpoint of 1.2 mm to be the most efficient cutpoint for sentinel lymph node biopsy result (p = 0.0004). Completion lymphadenectomy was performed in 46 sentinel node-positive patients; 12 (26.1 percent) had at least one additional positive nonsentinel node. Nonsentinel node positivity was marginally associated with the presence of multiple positive sentinel nodes (p = 0.07). At mean follow-up of 13.8 months, four of 241 sentinel node-negative basins demonstrated same-basin recurrence (1.7 percent). Sentinel lymph node biopsy is highly reliable in experienced hands but is a low-yield procedure in most thin melanomas. Patients with melanomas thicker than 1.2 mm or with ulcerated or high mitotic index lesions are most likely to have occult lymph node metastases by sentinel lymph node biopsy. Completion therapeutic lymphadenectomy is recommended after positive biopsy because it is difficult to predict the presence of positive nonsentinel nodes.     

DNA ploidy, Ki-67 and p53 as indicators of lymph node metastasis in early gastric carcinoma

OBJECTIVE: To perform flow cytometric and immunohistochemical analysis of early gastric carcinoma in order to investigate the clinical value of DNA ploidy, Ki-67 and p53 as indicators of lymph node metastasis. STUDY DESIGN: We studied 108 cases of surgically resected early gastric carcinoma, which comprised 21 cases showing positive lymph node metastasis (group A) and 87 cases showing negative lymph node metastasis (group B). RESULTS: Aneuploidy was seen in 16 cases (76.2%) in group A in contrast to 10 cases (11.5%) in group B (P = .0029). The mean value of the Ki-67 labeling rate in all cases was 21.9%, and 15 cases (79.0%) showed a Ki-67 labeling rate of more than 21.9% in group A, while they numbered 31 (37.4%) in group B (P = .0016). Cases with cell positive for p53 protein numbered 15 (75%) in group A, while they numbered 38 (46%) in group B (P = .0460). CONCLUSION: DNA aneuploidy, a high labeling rate of Ki-67 and overexpression of p53 are all useful indicators of lymph node metastasis in patients with early gastric carcinoma.     

The diagnostic value of thyroglobulin concentration in fine-needle aspiration of the cervical lymph nodes in patients with differentiated thyroid cancer

INTRODUCTION: Recurrent differentiated thyroid cancer generally occurs first in the neck. Ultrasound is sensitive in detecting enlarged cervical lymph nodes but is not specific enough. Ultrasound-guided fine-needle biopsy increases the specificity but still may fail to detect a recurrence of the disease in the cystic metastatic lymph nodes. The aim of the study was to estimate the value of Tg concentration in the needle washout after fine-needle aspiration of suspicious lymph nodes. MATERIAL AND METHODS: The 105 patients studied had presented one or more enlarged suspicious cervical lymph nodes. All had undergone total thyroidectomy and (131)I ablative therapy. Serum thyroglobulin (Tg) concentration was within the 0.15-711.5 ng/ml range (mean 22.24 ng/ml) and Tg recovery range 94-100%. The positive Tg washout concentration cut-off value was established as equal to the mean plus two standard deviations of the Tg washout concentration of patients with negative cytology. RESULTS: Lymph node involvement was diagnosed by cytology in 15 patients and in 28 lymph nodes. Positive Tg washout concentration was found in 22 patients and in 48 lymph nodes. All the lymph nodes which turned out to have positive cytology had a positive Tg washout concentration. All lymph nodes with positive cytology were positive in pathology. Seven patients and 20 lymph nodes with negative cytology were positive in the Tg washout concentration test. All but one patients and all but two lymph nodes with a positive Tg washout concentration had positive pathology. CONCLUSIONS: 1. Ultrasound-guided fine-needle biopsy is not sensitive enough to detect all metastatic lymph nodes. 2. The Tg washout concentration test is 100% sensitive in the detection of metastatic lymph nodes. 3. Cytology in ultrasound- guided fine-needle biopsy is 100% specific. 4. The Tg washout concentration test carries a risk of false-positive results. 5. Both methods should be used for early detection of metastatic lymph nodes in patients with differentiated thyroid cancer.