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 correlations of plasma D-dimer and fibrinogen levels with carcinoembryonic antigen (CEA) and cytokeratin 19 fragment (CYFRA21-1) in patients with non-small cell lung cancer (NSCLC). Methods The clinical data of 196 patients with NSCLC diagnosed for the first time in the Department of Respiratory and Critical Care Medicine, the 416 Hospital of Nuclear Indusry between July 2017 and December 2019 were analyzed retrospectively. There were 57 cases in early stage (stage Ⅰ-Ⅱ), 57 cases in medium stage (stage Ⅲ), and 82 cases in advanced stage (stage Ⅳ) according to TNM staging, 108 cases of adenocarcinoma, 87 cases of squamous cell carcinoma, and 1 case of unclassified type according to pathological classification, and 19 deaths and 177 survivals according to outcome. The levels of D-dimer and fibrinogen were determined by immunoturbidimetry and coagulation method, and the levels of CEA and CFYRA21-1 were determined by electro-chemiluminescence method. The non-normally distributed data were presented as median (lower quartile, upper quartile), and Spearman correlation analyses were performed. Results Among the early, middle and advanced stage patients, the levels of D-dimer [198.00 (133.00, 390.87), 279.00 (170.93, 520.89), 389.00 (196.25, 931.00) μg/L], CEA [3.20 (2.60, 5.17), 13.53 (5.07, 70.63), 15.69 (4.07, 123.46) μg/L], and CFYRA21-1 [4.79 (3.15, 8.84), 8.60 (4.83, 19.32), 7.19 (3.09, 15.05) μg/L] were significantly different (P<0.05); however, there was no statistical difference in the level of fibrinogen among the three stages (P>0.05). The level of CYFRA21-1 in the adenocarcinoma group was lower than that in the squamous cell carcinoma group [(5.39 (2.81, 12.71) vs. 6.86 (4.18, 12.29) μg/L, P<0.05], while there was no statistically significant difference in D-dimer, CEA, or fibrinogen between the two groups (P>0.05). The levels of D-dimer, CEA, and CFYRA21-1 in the death group [1176.00 (382.00, 2848.00), 135.34 (24.85, 403.50), 10.82 (7.41, 23.41) μg/L] were significantly higher than those in the survival group [270.00 (146.00, 481.50), 5.62 (3.05, 26.53), 6.28 (3.37, 12.30) μg/L], and the differences were statistically significant (P<0.01); but there was no statistical difference in the level of fibrinogen between the two groups (P>0.05). D-dimer was positively correlated with CEA and CFYRA21-1 (rs=0.450, 0.291; P<0.001), but fibrinogen was not correlated with CEA or CFYRA21-1 (P>0.05). Conclusion D-dimer was more valuable than fibrinogen in predicting the clinical stage and prognosis of NSCLC.
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 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).
【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.
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.
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 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 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.