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 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.
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.
【Abstract】Objective To investigate the detection and significance of micrometastasis in lymph nodes of periampullary carcinoma. MethodsThe immunoreactivity of CK19, CK7 and CK18 in 220 lymph nodes from 60 patients who had been carried out radical resection of periampullary carcinoma were revealed by immunohistochemical method. Combining with clinical data and the followup result discussion and analysis were made. All the lymph nodes collected from January 1997 to August 2004 in this hospital were examined to be nonlymphonodus metastasis by histological HE stain. The criterion of positive CK was established and micrometastasis will be diagnosed if CKs revealed to be positive combining with the morphological changes of lymph node tissue through the observation of microscope. ResultsFortythree of 220 lymph nodes of periampullary carcinoma had been micro metastasized. The detection rates of micrometastasis were 19.55%（43/220）， 14.55%（32/220）and 11.36%（25/220） with CK19，CK7 and CK18 antibody, respectively, which meant that CK19 were better than CK18 for detection of micrometastasis. Detection rates of CK7， CK19 were higher in Ⅲ， Ⅳ stage than inⅠ， Ⅱ（P＜0.05）, whereas it had no relationship with gender, age and tumor differentiation degree. The oneyear survival rate for micrometastasis patients was lower than nonmicrometastasis patients’ (P＜0.05). Conclusion Micrometastasis of periampullary carcinoma may be the early stage of tumor metastasia. The CK antibodies are significant levels to detect micrometastasis of periampullary carcinoma and to guide the clinical treatment and prognostic judgment.
Objective To identify micrometastasis in regional lymph nodes of gastric cancer by quantitative real-time reverse transcription-PCR (qRT-PCR) assay and to evaluate the clinical significance of micrometastasis. Methods To study 320 lymph nodes collected from January 2010 to June 2010， 281 of which were from 40 patients with gastric cancer who had undergone a standard gastrectomy with lymphadenectomy， and other 39 of which were from 10 patients with gastroduodenal ulcer. Made CEA， CK-19， and CK-20 as primers， and used qRT-PCR assay in addition to hematoxylin and eosin staining to detect the micrometastasis， and to analyze the clinicopathologic characteristics.Results Totally， micrometastasis were detected by qRT-PCR assay in 31 (15.34%，31/202) lymph nodes of 28 (70.00%， 28/40) patients. Thirty-nine lymph nodes from 10 patients with gastroduodenal ulcer were negative by qRT-PCR and HE staining. The degree of differentiation， depth of gastric mural invasion， and clinical stage had statistically significant correlation with the incidence of lymph node micrometastasis (P＜0.05). Conclusions qRT-PCR assay is a sensitive and specific method to detect lymph node micrometastasis in gastric cancer patients，and it has importantly clinical significance in evaluating clinical staging，prognosis and treatment prescription.
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.