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.
Abstract: Objective To evaluate the significance of expression of vascular endothelial growth factor-C （VEGF-C） and cytokeratin 19 （CK19） in patients with stage I non-small cell lung cancer （NSCLC）. Methods A total of 269 patients with NSCLC who underwent standard lobectomy and lymph node dissection by the same surgical team in our hospital from January 2004 to June 2005 were included in this study. All the clinical data and follow-up results were complete, and all the pathological specimens were well kept. No preoperative or postoperative adjuvant therapy such as radiotherapy and chemotherapy was administered to those patients. Expressions of VEGF-C in cancer tissues was detected by immunohistochemical streptavidin-peroxidase （S-P） method, and CK19 was marked to examine micrometastasis in hilar and mediastinal lymph nodes. Clinical outcomes, pathological results and follow-up data were analyzed in combination with VEGF-C and CK19 expression. Results VEGF-C expression was not statistically different between different category in sex（Hc=1.722，P=0.084）, age （Hc=0.914，P=0.360）, smoking （Hc=2.440，P=0.295）, pathology type （Hc=5.668，P=0.058）or tumor size （Hc=0.165，P=0.920） . VEGF-C expression was statistically different between different groups of pathological differentiation （Hc=29.178，P=0.000). CK19 expression was not statistically different between different category in sex（χ2=0.000，P=0.999）, age （χ2=0.005，P=0.999）, smoking （χ2=2.294，P=0.317）, pathology type （χ2=0.573，P=0.289）, tumor size（χ2=0.006，P=0.999）, and pathological differentiation （χ2=2.927，P=0.231）. Five-year survival rate was statistically different between different grade of VEGF-C expression （χ2=37.318，P=0.000), and was also statistically different between positive group and negative group of CK19 （χ2=39.987，P=0.000). There was statistical difference between different grade of VEGF-C expression and positive rate of CK19 （χ2=25.954，P=0.000). Conclusion Expression of VEGF-C and CK19 is closely related to postoperative 5-year survival of patients with stage I NSCLC. Detection of VEGF-C and CK19 is of great clinical significance as it is helpful to predict patient prognosis and choose proper postoperative adjuvant therapy.
OBJECTIVE: To investigate the selection and identification of human keratinocyte stem cells(KSC) in vitro. METHODS: According to the characteristics of KSC which can adhere to extracellular matrix very fast, we selected 3 groups of different time(5 minutes, 20 minutes and 60 minutes) and unselected as control group. And the cells were identified by monoclone antibody of beta 1-integrin and cytokeratin 19 (Ck19), then the image analysis was done. Furthermore we analyzed the cultured cells with flow cytometer(FCM) and observed the ultrastructure of the cell by transmission electron microscope(TEM). RESULTS: The cell clones formed in all groups after 10 to 14 days, while the cells of 5 minute group grew more slowly than those of the other groups, however, the clones of this group were bigger. The expression of beta 1-integrin and Ck19 were found in all groups. The positive rate of beta 1-integrin was significant difference between 5 minute group and the other groups (P lt; 0.05). And the expression of Ck19 was no significant difference between 5 minute group and 20 minute group(P gt; 0.05), and between 60 minute group and control group. But significant difference was observed between the former and the later groups(P lt; 0.05). The result of FCM showed that most cells of the 5 minute group lied in G1 period of cell cycle, which was different from those of the other groups. At the same time, the cells of 5 minute group were smaller and contained fewer organelles than those of the other groups. CONCLUSION: The above results demonstrate that the cells of 5 minute group have a slow cell cycle, characteristics of immaturity, and behaving like clonogenic cells in vitro. The cells have the general anticipated properties for KSC. So the KSC can be selected by rapid attachment to extracellular matrix and identified by monoclone antibody of beta 1-integrin and Ck19.
Objective To explore the expressions of galectin-3 （Gal-3） and cytokeratin-19 （CK-19） in different tissues of Hashimoto thyroiditis （HT） complicated with papillary thyroid microcarcinoma （PTMC）. Methods The tumor tissue， 0.5 cm near tumor tissue， and opposite lateral lobe thyroid tissue in 25 HT with benign nodus patients， 25 PTMC patients， and 25 HT with PTMC patients were collected. The expressions of Gal-3 and CK-19 in these tissues were detected by immunohistochemical methods. Results ①The positive rates of Gal-3 and CK-19 expressions in the tumor tissueof HT with PTMC patients and PTMC patients were significantly higher than those of HT with benign nodus patients （P＜0.05）.②The positive rates of Gal-3 and CK-19 expressions in the opposite lateral lobe thyroid tissue of HT with PTMC patients and HT with benign nodus patients were significantly higher than those of PTMC patients （P＜0.05）．③The positive rates of Gal-3 and CK-19 expressions in the 0.5 cm near tumor tissue of HT with PTMC patients and HT with benign nodus patients were significantly higher than those of PTMC patients （P＜0.05）. ④The middle and b positive rates of Gal-3 and CK-19 expressions in the 0.5 cm near tumor tissue of HT with PTMC patients were significantly higher than those of HT with benign nodus patients and the PTMC patients （P＜0.05）．Conclusions ①Gal-3 and CK-19 protein are helpful to differentiate the benign thyroid tumor and malignant one. ② The expressions of Gal-3 and CK-19 protein in patients with HT are clear higher than those in patients without HT that means the prognosis evaluation in HT canceration. ③ Gal-3 combined with CK-19 protein are help for early diagnosis， the pathogenesis and prognosis evaluation in thyroid cancer. The b positive means canceration. ④ In HT with PTMC， it needs an operation therapy and a larger one， which is appropriate for lateral and opposite lobe partial resection or total resection.
Objective To analyze the expressions of galectin-3， human bone marrow endothelial cell-1 （HBME-1），cytokeratin （CK）19， and RET in benign and malignant thyroid tumor and to discuss their clinical significances. Methods The clinicopathologic and immunohistochemical staining data of 131 patients with benign and malignant thyroid tumor were analyzed retrospectively， including 45 patients with malignant thyroid tumor， 86 patients with benign thyroidtumor. The expressions of galectin-3， HBME-1， CK19， and RET in the benign and malignant thyroid tumor were detectedby immunohistochemical staining. Results The positive expression rates of the galectin-3， HBME-1， CK19， and RET in the malignant thyroid tumor were 97.8% （44/45）， 88.9% （40/45）， 100% （45/45）， and 71.1% （32/45）， respectively，which in the benign thyroid tumor were 9.3% （8/86）， 12.8% （11/86）， 37.2% （32/86）， and 8.1% （7/86）， respectively， the differences were statistically significant （P＜0.05）. The diagnostic sensitivity， specificity， and accordance rates were 97.8 %， 90.7%， and 93.1% for the galectin-3， respectively；88.9%， 87.2%， and 87.8% for the HBME-1， respec-tively；100%， 62.8%， and 75.6% for the CK19， respectively；71.1%， 91.9%， and 84.7% for the RET， respectively. Conclusions The expression levels of galectin-3， HBME-1， CK19， and RET in malignant thyroid tumor are significantly higher than those in benign thyroid tumor. Galectin-3， HBME-1， CK19， and RET can be important factors for identifying the benign and malignant tumor and their biological behaviors. Galectin-3 has a high reference value in the diagnosis of thyroid carcinoma.
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.
【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.