Objective To investigate the clinical meanings of lymphangiogenesis, lymph vessel invasion (LVI) and lymph node (LN) micrometastasis in gastric cancer. Methods The expression of D2-40 in 68 patients with gastric cancer of primary lesion and the expressions of CK20 and (or) CKpan in 791 lymph nodes from 51 cases which were detected by immunohistochemical staining were analyzed, as well as their clinicopathologic profiles. The relationship of lymph vessel density (LVD), LVI and LN micrometastasis with LN metastasis and other clinicopathologic parameters was analyzed respectively. Results Positive rate of LVI with HE (LVI-HE) and D2-40 (LVI-IM) staining was respectively 66.2%(45/68) and 76.5%(52/68), P=0.118. The positive rate of LVI-IM was related to deeper tumor invasion (P=0.044), later stage of TNM (P=0.003) and LN metastasis (P=0.000). Average LVD of 68 cases was (18.19±7.44)/HP. The increment of LVD was significantly associated with LVI-HE positive status （P=0.040）, LVI-IM positive status （P=0.001）, venous invasion （P=0.037）, later stage of TNM （P=0.020） and LN metastasis （P=0.001）. The survival rate of the group sharing ≥15/HP of LVD was significantly lower than that in the group sharing ≤14/HP of LVD in early period of follow-up （P=0.032）. The incidence of nodal involvement in 51 patients was increased from 74.5%(38/51) by HE staining to 88.2%(45/51) by CK (CK20 or CKpan) immunostaining. The detection rate of metastasized LN was increased from 32.0%（253/791） by HE staining to 41.5%（328/791） by CK immunostaining (Plt;0.001). The significant difference of LN micrometastasis detection rate between CK20 (8.7%) and CKpan (12.3%) was also identified (P=0.003). The increased number of LN micrometastasis was related to larger diameter of tumor (P=0.001), higher LVI-HE positive rate (P=0.040), deeper invasion of tumor (P=0.018) and later stage of TNM (P=0.012). Both LN stage and TNM stage were changed according to the detection of LN micrometastasis: Seven patients of N0 should be recognized as N1 (N0→N1)， 6 as N1→N2, 1 as N2→N3. Four patients of stage Ⅰb should be recognized as stage Ⅱ (Ⅰb→Ⅱ)， 4 as Ⅱ→Ⅲa， 3 as Ⅲa→Ⅲb， 1 as Ⅲb→Ⅳ. Conclusion Detection of D2-40 and CK in diagnosis of LVI and LN micrometastasis is better than HE staining. The combined detection of CK20 and CKpan may be much easier to find out the LN with micrometastasis. Later stage of TNM the tumor is, more frequently the LN micrometastasis happens. The relationships of LVI-IM, LVD and LN micrometastasis with LN metastasis in gastric cancer has been demonstrated. Patients with higher LVD share a lower survival rate in early period of follow-up.
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 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.
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.
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 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.