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.
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.
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 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 explore the risk factors the central cervical lymph node micrometastasis of papillary thyroid microcarcinoma (PTMC). Methods PTMC patients who underwent surgical operations in West China Hospital, Sichuan University between January 2014 and December 2018 were retrospectively enrolled. The patient did not find lymph node metastasis in the central cervical area by preoperative ultrasound. During the operation, the central cervical lymph node of the affected side was dissected or lymph node dissection in the central area of the affected side of the neck plus the lateral area of the neck. With postoperative pathology as the gold standard, patients were divided into central cervical lymph node micrometastasis group (micrometastasis group) and central cervical lymph node non-metastasis group (non-metastasis group). The differences of clinical features and ultrasonic signs between the two groups were analyzed. Results A total of 507 patients were included, including 223 (44.0%) in the micrometastasis group and 284(56.0%) in the non-metastasis group. The results of univariate analysis showed that compared with the non-metastasis group, the patients in the micrometastasis group were younger, the tumor size were higher, the proportion of male, multifocality, bilateral involvement and thyroid capsular invasion were higher. The results of multiple logistic regression analysis showed that lower age [odds radio (OR)=0.967, 95% confidence interval (CI)(0.949, 0.985), P<0.001], male [OR=2.357, 95%CI (1.503, 3.694), P<0.001)], a larger maximum diameter of PTMC [OR=1.232, 95%CI (1.100, 1.379), P<0.001], a larger nodule volume of PTMC [OR=1.031, 95%CI (1.008, 1.114), P=0.032], multifocal lesion [OR=2.309, 95%CI (1.167, 4.570), P=0.016] and invasion of the thyroid capsule [OR=1.520, 95%CI (1.010, 2.286), P=0.045] were independent risk factors for central cervical lymph node micrometastasis. Conclusions The patient’s male, young age, PTMC nodule with large maximum diameter and large volume, multifocal, and invasion of the thyroid membrane are risk factors for the central cervical lymph node micrometastasis of PMTC patients. These clinical and ultrasound signs can provide a theoretical basis for doctors’ clinical management decisions.
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 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 determine the value of detection of micrometastasis in peripheral blood to hepatocellular carcinoma （HCC） metastasis or recurrence． Methods Reviewed the related literatures, the methods and significances of the detection of HCC micrometastasis in peripheral blood were analyzed. Results Currently, there are mainly two methods, hematogenous dissemination cell detection and HCC specific mRNA biomarker detection, for detection of HCC micrometastasis in peripheral blood. Theoretically, although they are considered as early detections of HCC metastasis or recurrence, researches still not have a abroad agreeable conclusion from different studies. After adjusting and improving the methods and detection time, different studies also have not gotten a quite consistent conclusion. Conclusion There is a great significance in detection of HCC micrometastasis in peripheral blood to understanding the mechanisms of HCC metatasis and recurrence, and also to improving the clinical therapy. Theoretically and practically, the method should be improved for facilitating the mechanism research of HCC metastasis and recurrence, and the application of detection.
Abstract： Objective To evaluate the sensitivity, specificity and clinical significance of Lunx mRNA in surveying micrometastasis by sampling peripheral blood of lung cancer patients, studying the early diagnosis of lung cancer metastasis. Methods From March 2004 to February 2005，Reverse transcriptionpolymerase chain reaction(RT-PCR) was used to detect Lunx mRNA of peripheral blood of 60 lung cancer patients（lung ancer group）. Peripheral blood of 20 patients with pulmonary benign lesions (pulmonary benign lesions group) and 10 normal healthy volunteers (control group) were used as control. Results (1) In the lung cancer group, Lunx mRNA were expressed positive in 28(46.7％) patients. All the pulmonary benign lesions group (0/20) and the control group (0/10) were expressed negative. (2) One of the 12 stage I patients with lung cancer (8.3％) was positive for Lunx mRNA, 5 of the 15 stage Ⅱ patients (33.3%) were positive, 22 of the 33 stage Ⅲ patients (66.7%) were positive. Comparing the positive rate of these groups, there was no statistically difference between stage Ⅰ and stage Ⅱ, but the difference between stage Ⅰ+ stage Ⅱ and stage Ⅲ significant (χ2=15.88, P=0.000). (3) In 38 adenocarcinoma, 17 were positive for Lunx mRNA. In 14 squamous carcinoma, 7 were positive. All the 3 adenosquamous carcinoma expressed positive. 1 of 3 small cell lung cancer was positive, 1 large cell carcinoma and 1 carcinoma sarcomatodes expressed negative. Comparing the positive rate of these groups, there was no statistically difference among them. (4) By followup till March 2005, 10 lung cancer patients were found metastasis. Among them, 9 were positive for Lunx mRNA expression, and 1 was negative. Conclusion Lunx mRNA has high sensitivity and specificity in surveying micrometastasis by ampling peripheral blood. It would likely to be an proper gene for the detection of micrometastasis in lung cancer patients.