Objective To investigate the lymph node micrometastasis and its clinicopathologic features on 5-year disease free survival rate for patients with pT1—3N0 gastric cancer. Methods One hundred and twenty patients with stage pT1—3N0 gastric tumors were included, and 2 106 lymph nodes were harvested and examined in all the specimens. There were 9－28 lymph nodes with average 18 lymph nodes from each patient. All the lymph nodes were negative by HE staining. The CK20 expression of lymph nodes was tested by immunohistochemistry. The relationships between clinicopathologic features or CK positive expression and 5-year disease free survival were analyzed. Results The positive expression rate of CK20 was 9.07% (191/2 106) in lymph nodes and 26.67% (32/120) in patients with pT1—3N0 gastric cancer by immunohistochemistry. Eleven cases were with micriometastasis, 21 cases were isolated tumor cells (ITC). The average postoperative follow-up was 66.35 (range 24—121) months. Five-year disease free survival rates were 87.4%, 78.3%, and 40.9% for the lymph node negative, ITC, and micrometastasis groups, respectively. Five-year disease free survival rate in the micrometastasis group was lower than that in the lymph node negative group (P=0.000) and ITC group (P=0.046). However, there was no significant difference between the lymph node negative group and ITC group （P=0.253）. Multivariate analysis identified tumor diameter (P=0.011), depth of tumor invasion (P=0.043), and lymphatic vessel invasion (P=0.002) were related with CK20 positive expression. There was no significant relationship between the pathologic parameters and the 5-year disease free survival rates. Lymph node micrometastasis of gastric cancer was detected in 11 patients who should belong to stage pN1(Mi), the restage rate was 9.17%. While the lymph node negative (88 patients) and ITC (21 patients) were recorded pN0(i-) and pN0(i+), respectively, and were not recommended restage (stage pN0). Conclusion Patients with stage pT1—3N0 gastric cancer and micrometastasis in lymph node are with high-risk and low 5-year disease free survival rate, for whom adjuvant therapies may be justified and effective.
Objective To reveal the significance of D2-40/CK19 dual immunohistochemistry for micrometastasis of peripancreatic neural plexus in patients with pancreatic cancer. Methods Between January 2006 and January 2007, 44 patients with pancreatic duct adenocarcinoma underwent extended radical resection. Conventional hematoxylin/eosin staining and double immunohistochemical staining using CK19 and D2-40 were used to determine peripancreatic neural invasion and lymphatic vessel invasion (LVI) in peripancreatic neural plexus tissues. Results D2-40 immunohistochemistry showed brown-yellow tube-like lymph vessels. The lymph vessel of peripancreatic nerve plexus followed vascular and perineurium, and the lymph vessel adjacent to peripheral nerve fascicles owned tube-like structure. CK19 immunohistochemistry showed cytoplasm of pancreatic cancer cell was red. The LVI was observed in lymphatic capillaries. Peripancreatic neural plexus invasion was found in 30 cases (68.2%), tumor cell invading presented in lymph vessels of peripancreatic neural plexus in 21 patients (47.7%) with pancreatic cancer. The peripancreatic neural plexus invasion was associated with LVI (P=0.003). The plexus of pancreatic capitalis and celiac plexus were respectively confirmed to be the spot with the highest lymphatic vessel density and the maximal incidence of neural plexus invasion simultaneously. Conclusions Patients with pancreatic cancer should be given the opportunity of radical operation combining related peripancreatic neural plexus as far as possible. The dual immunohistochemical staining with anti-CK19 and anti-D2-40 monoclonal antibodies should be a new method in research of perineural invasion of pancreatic cancer, exhibiting both the pancreatic cancer cells and lymph vessels clearly and distinctly.
Objective To detect the expression of cytokeratin 20 (CK20) mRNA (micrometastasis) in regional lymph nodes and the serum activities of CD4+ cells, CD8+ cells and NK cells, serum levels of IL-2, IL-12 and sIL-2R in peripheral blood of patients with colorectal cancer; and to investigate the relationship between them. Methods Total 281 lymph nodes of 21 patients with colorectal cancer were collected. The positive expression of CK20 mRNA in lymph nodes was detect by reverse transcription-polymerase chain reaction (RT-PCR) and the metastasis in lymph nodes was detected by conventional pathological examination; the serum activities of CD4+ cells, CD8+ cells and NK cells were detected by flow cytometry and serum levels of IL-2, IL-12 and sIL-2R were detected by ELISA method in peripheral blood of patients with colorectal cancer. Results Among the positive metastasis in the 281 lymph nodes of the 21 patients, there were 16 (5.7%， 16/281) lymph nodes in 2 patients detected by pathological examination and 140 (49.8%， 140/281) lymph nodes in 10 patients by RT-PCR. There was a significant difference between the two measures in the aspects of the detection rate and the positive cases of lymph node metastasis in the 21 patients. Before operation, the serum activities of CD4+ cells, CD4+/CD8+ and NK cells, levels of IL-2 and IL-12 in 11 patients whose CK20 mRNA in regional lymph nodes were negative expression were higher than those in the other 8 patients whose lymph nodes metastasis were negative by conventional pathological examination but CK20 mRNA were positive expression （P＜0.05）; and the serum activity of CD8+ cells and level of sIL-2R in the former ones were lower than those in the latter ones （P＜0.05）. The serum activities of CD4+（r＝－0.769） cells, CD4+/CD8+（r=－0.755） and NK cells （r=－0.532）, the levels of IL-2 （r＝－0.834） and IL-12 （r＝－0.819） were negative correlated with the expression of CK20 mRNA （P＜0.05, P＜0.01）; and the activity of CD8+ cells （r=0.562） and level of sIL-2R （r=0.751） were positive correlated with the expression of CK20 mRNA （P＜0.05）. Conclusion The micrometastasis in lymph nodes is correlated significantly with the lower immune function of patients with colorectal cancer.
Objective To find and evaluate the existence of distant peritoneal micrometastasis of gastric cancer in rectovesical pouch or Douglas pouch by using immunohistochemist ry method. Methods Forty cases of gastric cancer were collected f rom June 2004 to March 2006 in Nanjing Gulou hospital . None of them showed obvious distant peritoneal metastasis in preoperative physical and imaging examinations and laparotomy inspection or palpation. Tissues were taken f rom rectovesical pouch or Douglas pouch during the operations , and HE and CEA/ CK220 immunohistochemistry staining were then performed on the tissues. Results Distant peritoneal micrometastasis in rectovesical pouch or Douglas pouch were found in 10 cases out of the 40 cases , all of which were found to have full-thickness invasion or invasion out side gast ric serous tunic 〔27. 8 % (10/ 36) 〕. Their occurrence rates of peritoneal micrometastasis were significantly higher than those without full-thickness invasion〔0 (0/ 4) 〕, Plt;0. 05. The number of metastatic lymph nodes was more than six in 8 cases , was only one in 2 case , the occurrence rate of peritoneal micrometastasis of the number of metastatic lymph nodes was more than seven 〔44. 4 %(8/ 18) 〕which was significantly higher than that the number was less than seven〔16. 7 % (2/ 12) 〕, Plt;0. 05. In 10 cases , 8 cases were poorly differentiated adenocarcinoma , and the other two were moderately differentiated. Conclusion When gast ric carcinoma invaded serous tunic or outside , though peritoneal metastasis may not be found by preoperational inspection or intraoperative palpation , peritoneal biopsy in rectovesical pouch or Douglas pouch may be necessary to perform as a routine procedure to detect distant peritoneal micrometastasis. It may be useful for staging , adjuvant chemotherapy and prognosis forecast.
ObjectiveTo investigate the clinical significance of CK20 mRNA expression in blood of patients with colorectal cancer. MethodsThe expressions of CK20 mRNA in blood of twenty healthy volunteers, ten patients with colorectal polyp and sixtyone patients with colorectal cancer were detected by RT-PCR. ResultsThe positive rate of CK20 mRNA in peripheral venous blood and portal venous blood of patients with colorectal cancer were 41.0%(25/61) and 45.9%(28/61), which was not significantly different (Pgt;0.05). The expression of CK20 mRNA in patients with colorectal cancer was associated with clinical TNM stage of tumor, local lymph node metastasis, distance metastasis, and the depth of invasion (Plt;0.05). No expression of CK20 mRNA was detected in blood of twenty healthy volunteer’s and ten patients with colorectal polyp. ConclusionCK20 is a specific marker for detecting blood micrometastasis of colorectal cancer. The expression of CK20 mRNA in blood of patients with colorectal cancer is related with TNM stage, invasion, and metastasis of colorectal cancer.
Objective To study the significance of c-met mRNA in axillary drainage after operations for breast cancer. Methods RT-PCR assay was used to examine c-met mRNA in axillary drainage after operations in 52 cases of breast cancer. The relationships between the expression of c-met and the tumor size, metastatic lymph nodes， the expressions of estrogen receptor (ER), progesterone receptor (PR) and c-erbB-2 were analyzed, respectively. In addition, the effect of douching operative field with 5-FU and distilled water on the expression of c-met mRNA was also analyzed. Results ①The proto-oncogene c-met mRNA could be detected in axillary drainage after operations for breast cancer by RT-PCR, and its positive rate was higher than that in routine pathological detection for micrometastasis in the axillary lymph nodes (P＜0.05). ②The expression of c-met mRNA was correlated with both the metastatic lymph nodes and tumor size. ③There was no significant relationship between the expression of c-met mRNA and the expressions of ER, PR and c-erbB-2. ④Dounching operative field with 5-FU and distilled water could decrease the expression of c-met mRNA.Conclusion The proto-oncogene c-met mRNA may be an ideal and specific marker for dectecting micrometastasis of breast cancer. In addition, it also suggests that the examination of c-met mRNA in the axillary drainage by RT-PCR assay could detect the micrometastasis in axillary lymph nodes much easier and more accurately than routine pathological method.
ObjectiveTo study the detection methods of micrometastasis in sentinel lymph nodes (SLN) and their clinical significance. MethodsFifty women with breast carcinoma were included. SLN in fifty breast carcinoma was identified by using methylene blue staining to detect and remove them for routine hematoxylin and eosin stain and histological exam. All negative SLNs were examined by serial section (SS) with the section interval of 250 μm and HE stain for microscopic examination and immunohistochemical (IHC) exam was performed with CK19 monoclonal antibody. Then the above three detection methods were analyzed. All patients had axillary lymph node dissection (ALND)，and all none sentinel lymph nodes (NSLN) were examined by Hamp;E staining.ResultsThe SLNs were identified in 45 of 50 patients with a detection rate of 90%. Sixteen SLNs were found positive with routine histological exam, the positive detecting rate was 35.56%, while the other 29 negative SLNs were found 7 and 6 cases of micrometastasis using SS and IHC methods，therefore the positive detecting rate was increased by 15.55% and 13.33%, respectively.Conclusion SS and IHC methods could detect the micrometastasis in negative SLN with routinely histological exam, increasing the positive detecting rate and decreasing the false negative rate.
Objective This study aimed to investigate the predictive value of preoperative serum CA19-9 level for lymph node micrometastasis in patients with lymph node metastasis-negative gastric cancer and its effect on prognosis. Methods Clinicopathological data were retrospectively collected from 176 cases of gastric cancer who underwent D2 radical surgery in our hospital between January 2006 and December 2011, and also collected the patients’ lymph node tissue specimens. All patients were confirmed by pathologic examination of lymph node metastasis-negative. Quantitative real-time PCR (qRT-PCR) was used to detect the presence of lymph node micrometastasis in lymph node tissues. Sixty cases of gastric cancer were selected to construct the receiver operating characteristic curve (ROC) of preoperative serum CA19-9 level to predict lymph node micrometastasis, then established the threshold value. The remaining 116 cases were used to validate the rationality of this threshold. In addition, we explored the impact of preoperative serum CA19-9 level on the prognosis of patients with lymph node metastasis-negative gastric cancer, and explored the risk factors of lymph node micrometastasis. Results ① Results of ROC curve: the preoperative serum CA19-9 level of 15.5 U/mL was the threshold for predicting lymph node micrometastasis, with a sensitivity of 93.1%, specificity of 63.6%, and area under the curve (AUC) of 0.84 (P=0.003). With 15.5 U/mL as the threshold, 116 patients were divided into positive group and negative group. The lymph node micrometastasis rates in the 2 groups were different, which was higher in the positive group than that in the negative group (P<0.001). ② Effect of preoperative serum CA19-9 level on prognosis: the patients were divided into the positive group and the negative group with 15.5 U/mL as the threshold, and the log-rank test showed that the survival of the negative group was better than that of the positive group (P=0.001). ③ The risk factors for lymph node micrometastasis: the logistic regression model showed that preoperatively positive serum CA19-9 was an independent risk factor for lymph node micrometastasis in patients with gastric cancer [OR=1.860, 95% CI was (1.720, 2.343), P<0.001]. Conclusion Preoperative serum CA19-9 level can be used to predict lymph node micrometastasis in lymph node metastasis-negative patients with gastric cancer.