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 Epidermal stem cells (ESCs) can actively partici pate in wound heal ing and enhance reepithel ial ization. To establ ish ideal diabetes mell itus (DM) rat models and to investigate the expression of keratin 19 (K19),β1-integrin, β-catenin, and prol iferating cell nuclear antigen (PCNA) in ESCs of DM rat model, then to study the potential mechanism of difficult recovering wounds of diabetic skin. Methods Twenty male SD rats (weighing 250-300 g) were dividedinto DM group and normal control group randomly (n=10). The DM rat model was made by intraperitoneal injected 65 mg/kg streptozocin (STZ), the normal control group was not treated. At 4 weeks after injection, pancreatic tissue was harvested for HE staining in two groups. The ESCs isolated from full-thickness skins of the back of two group rats were culutured and identified. The 2nd passage of ESCs were obtained for immunocytochemical staining of K19, β1-integrin, β-catenin, and PCNA. Meanwhile, the cell cycle were measured by flow cytometry. The cell colony formation rates were detected after 1 week. Results The achievement ratio of DM rat model was 90% with good stabil ity. HE staining showed that the number of islet cells significantly decreased with degeneration and necrosis in DM group; the structure of islet cell was clear without degeneration and necrosis in normal control group. The integral absorbance values of positive expression for K19, β1-integrin, β-catenin, and PCNA in ESCs of DM group (82.63 ± 14.77, 21.59 ± 4.71, 6.49 ± 6.58, and 90.77 ± 12.44, respectively) were significantly lower than those in the normal control group (151.24 ± 42.83, 54.48 ± 17.43, 116.39 ± 9.26, and 110.62 ± 20.67, respectively) (P lt; 0.01). The clone forming efficiency of ESCs in DM group (6.43% ± 1.01% ) was significantly lower than that in the normal control group (11.37% ± 1.62%) (P lt; 0.01). Flow cytometry indicated that 88.89% of cultured ESCs in the DM group were in resting state/ pre-DNA-synthetic gap (G0/G1), and the apoptosis rate was 3.98%; 91.50% in the normal control group and the apoptosis rate was 0. Conclusion The DM rat model can be effectively induced by intraperitoneal injected 65 mg/ kg STZ. The decreased amount and the low prol iferation and differentiation capacity of ESCs may be one of the important mechanisms of difficult recovering wounds of DM rats.
Objective To investigate the effect of keratin 17 (K-17) on the migration, prol iferation and tube formation of human umbil ical vein endothel ial cell (HUVEC), and to real ize the role of K-17 in angiogenesis. Methods After HUVEC were cultured in DMEM medium supplemented with 10%FBS overnight, K-17-siRNA-mixture (experimental group) and Ncontrol-siRNA-mixture (negative control group) were added into HUVEC, respectively, by Lipofectamine 2000 transfection assay, and the final concentration of the siRNA was 50 nmol/L. Lipofectamine 2000 alone was used as the control. After the cells were cultured for 36 hours, the cell prol iferation abil ity was detected by cell counting. After 30-hour culture, the cell’s abil ities of migration and differentiation to tube were detected by 24-well Mill icell units and the collagen gel assay, respectively. In addition, non-siRNA-treated HUVEC were cultured for 24 hours in DMEM medium supplemented with 10%FBS (group A), 2%FBS (group B) and 2%FBS+10 ng/mL bFGF (group C), respectively, and then the expression of K-17 in HUVEC was detected by RT-PCR and Western blot. Results After the treatment with K-17-siRNA for 36 hours, HUVEC exhibited no significant difference in the prol iferation, compared with both control and negative control groups (P gt; 0.05). After transfected with K-17-siRNA for 30 hours, the number of HUVEC in the experimental group which migrated from the upper chamber to the lower chamber of Mill icell wells within 24 hours (3719.0 ± 319.0) was smaller than both control (7 437.5 ± 212.0) and negative control (7 356.3 ± 795.7) groups, with significant difference (P lt; 0.01). However, there was no significant difference between the control group and the negative control group (P gt; 0.05). After HUVEC were transfected with K-17- siRNA for 30 hours, the number of tubes in the experimental group, the negative control group and the control group in 24 hours was (1.1 ± 0.5), (3.6 ± 0.5) and (3.2 ± 0.6) per field, respectively. The experimental group was significantly different from both control and negative control groups (P lt; 0.01), and there was no significant difference between the negative control group and the control group (P gt; 0.05). The expression of K-17 protein in HUVEC in groups A, B and C was 0.25 ± 0.02, 0.08 ± 0.01 and 0.72 ± 0.03, respectively. There was significant difference among these three groups (P lt; 0.01). Conclusion K-17 has no impact on cell prol iferation, but may augment endothel ial cell migration, which may facil itate angiogenesis.
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 evaluate the diagnostic value of antikeratin antibody (AKA) for rheumatoid arthritis (RA). Methods Systematic and comprehensive literature was searched in PubMed (1966 to June 2010), The Cochrane Library (Issue 6, 2010), CBM (1978 to June 2010), CNKI (1994 to June 2010), VIP (1989 to June 2010), and CMA Digital Periodicals (1997 to June 2010). The diagnosis studies of antikeratin antibody for rheumatoid arthritis were included. The quality assessment of diagnostic accuracy studies (QUADAS) items were used to assess the quality of the included studies. The Meta-Disc (version 1.4) software was used to analyze the data. Results A total of 69 trials involving 14 890 participants were included. The results of meta-analyses showed that compared with the RA classification criteria revised by American Rheumatism Association (ARA), the summary sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, OR value, and summary receiver operating characteristic curve of antikeratin antibody were 0.41 (0.39, 0.42), 0.94 (0.94, 0.95), 9.52 (7.21, 12.57), 0.63 (0.60, 0.66), 15.24 (11.62, 19.98), and 0.613 6, respectively. Conclusion Antikeratin antibody might be one of the most effective diagnoses for rheumatoid arthritis. The clinicians should combine other autoantibodies with AKA to diagnose rheumatoid arthritis.
【摘要】 目的 探讨细胞角蛋白19的可溶性片段CYFRA21-1在慢性肾脏病（chronic kidney disease，CKD）患者血清中的表达及其临床意义。 方法 2008年10月-2009年4月随机选取45例CKD患者，根据肌酐清除率（creatinine clearance rate，Ccr）的大小分为Ccrlt;15 mL/min组（23例）和Ccrgt;15 mL/min组（22例），并以正常体检者（10例）为正常对照组，肺鳞癌患者（10例）为阳性对照组，每位参与研究者取血后采用酶联免疫吸附法测定血清CYFRA21-1的浓度。 结果 正常对照组、阳性对照组、Ccrgt;15 mL/min组和Ccrlt;15 mL/min组的血清CFRA21-1浓度分别为（1.720±0.535）、（21.010±11.809）、（3.310±1.569）和（5.090±1.306） ng/mL。与正常对照组比较，其余3组的CYFRA21-1浓度均显著升高，有统计学意义（Plt;0.05），且均值大于正常参考值（0~3.3 ng/mL）；Ccrgt;15 mL/min组和Ccrlt;15 mL/min组的CFRA21-1浓度显著低于阳性对照组，有统计学意义（Plt;0.05）；两个实验组间随着Ccr的降低，CYFRA21-1浓度升高，有统计学意义（Plt;0.05）。 结论 CKD患者血清CYFRA21-1水平的升高与肾功能减退存在一定关系，可将其作为临床上预测肾脏纤维化程度的指标。【Abstract】 Objective To discuss the clinical Between significance of cytokeratin 19 fragment (CYFRA21-1) in patients with chronic kidney disease (CKD). Methods From October 2008 to April 2009, 45 inpatients were randomly selected and assigned into three groups according to creatinine clearance rate (Ccr) level: Ccrlt;15 mL/min, Ccrgt;15 mL/min. Ten healthy volunteers were chosen as control group, and other 10 patients with lung squamous cell carcinoma as positive control group. ELISA was used to measure the serum concentration of CYFRA21-1. Results Compared with control group, the serum concentration of CYFRA21-1 in CKD groups and positive control group was elevated (Plt;0.05). As the Ccr decreased, the serum concentration of CYFRA21-1 was elevated (Plt;0.05) in two CKD groups. Conclusion Serum concentration of CYFRA21-1 in CKD patients has a relationship with the renal function decrease, and may be used as an indication of renal interstitial fibrosis (RIF).
目的:观察和探讨细胞角质素CK18慢性胆囊炎患者的胆囊组织和血清中的表达及其意义。方法:35例经腹腔镜胆囊切除的慢性结石性胆囊炎患者(27例女性患者,8例男性患者,年龄在55.65±13.48岁),将患者分为两个组,A组为患慢性非活动性结石性胆囊炎者(n=10),B组为患慢性活动性胆囊炎者(n=25),在细胞凋亡早期胱门蛋白酶分裂的CK18用M30细胞凋亡酶联免疫吸附测定,总细胞角蛋白18(从凋亡及坏死细胞中分离)用M65酶联免疫吸附测定。然后计算M30/M65结果:胱门蛋白酶分裂的CK18,特别是总CK18在胆汁中的表达远高于血清。在B组中,胱门蛋白酶分裂的CK18和总CK18的表达在胆囊组织和血清中表达差异相当大。在胆囊粘膜上皮细胞胱门蛋白酶分裂的CK18染色呈强阳性。结论:CK18在胆囊上皮细胞中表达。胱门蛋白酶分裂的CK18和总CK18在胆囊组织中的表达远高于血清中的表达。胱门蛋白酶分裂的CK18和总CK18的表达水平在活动性胆囊炎和非活动性胆囊炎中的表达并无明显差异。
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.