【Abstract】Objective To study the relationship of the expression of CD44v6 mRNA and nm23H1 mRNA with the clinical pathology parameter and prognosis of breast cancer, and to investigate the correlation of the expression of CD44v6 mRNA and nm23H1 mRNA. Methods In situ hybridization and CSA immunohistochemistry method were used to detect the expression of CD44v6 mRNA and nm23H1 mRNA in 94 cases of breast cancer. Results The positive expression of CD44v6 mRNA and the negative expression of nm23H1 mRNA were positively correlated with the grading, clinical stage, lymph node metastasis, and recurrence and prognosis of breast cancer. CD44v6 mRNA expression and nm23H1 mRNA were negatively correlated in breast cancer. Patients who had positive expression of CD44v6 mRNA and negative expression of nm23H1 mRNA had a higher lymph node metastatic rate and a lower survival rate. Conclusion Several genes were involved in the occurrence and development of breast cancer in which the expression of CD44v6 mRNA has synergistic action in negative regulation with that of nm23H1 mRNA. Combined detection of the expression of these two mRNA is helpful to judge the metastasis, recurrence and prognosis of breast cancer.
【Abstract】ObjectiveTo study the relationship between expression of CD44v6 in gastric carcinoma and neoplasm metastasis and prognosis. MethodsExpression of CD44v6 in 52 cases of gastric carcinoma was assayed by flow immunocytometry, and its relation with clinical pathology and prognosis was analyzed. ResultsIn 52 cases of gastric carcinoma tissue, the positive rate of CD44v6 expression was 67.31%(35/52); but the positive rate of CD44v6 expression in normal gastric tissue was 25.00% (13/52). The positive rate of expression was significantly different (P＜0.01). The positive rate of CD44v6 expression in gastric carcinoma tissues was related to the depth of carcinoma infiltration, lymph node metastasis and pTNM stage （P＜0.05）. ConclusionExpression of CD44v6 plays an important role in invasion, metastasis and pTNM stage of gastric carcinoma. It may be used as a new indicator to predict metastatic potential and prognosis of gastric carcinoma.
Objective To investigate the percentage of CD4 + CD25 + Treg cells and expression of Foxp3 mRNA in asthmatic patients and the impacts of inhaled steroids.Methods The percentages of CD4 +CD25 + Treg cells was assayed by flow cytometry and the expression of Foxp3 mRNA was detected by RT-PCR in peripheral blood mononuclear cells from the patients with chronic persistent asthma before and after steroids inhalation in comparison with healthy control. The forced expired volumin one second/predicted value( FEV1% pred) and peak expired flow( PEF) were measured by spirometry. Results The level of CD4 + CD25 + Treg cells and the expression of Foxp3 mRNA were lower in asthmatics before steroids treatment than those in control ( P lt; 0. 05) which were increased significantly after steroids treatment ( P lt; 0. 05) .FEV1% pred and PEF were declined significantly than those in control but improved markedly after treatment ( P lt; 0. 05) . Conclusions The insufficiency of amount and function of immue-suppressive CD4 + CD25 +Treg cells may play a role in the pathogenesis of asthma. Inhaled steroids can improve the lung function of asthmatics by upregulating the level of CD4 + CD25 + Treg and Foxp3.
To diminish the specific lymphocytes that responsive to the rejection of allograft. Anti-rat CD4,CD8 monoclonal antibodies and trichosnthin (TCS) was conjugated to immunotoxin by heterobifunctional reagent SPDP, 2-IT. The free TCS was removed from conjugates mixture by a column of Sephacryl S-200. The SDS-PAGE and cytotoxic assay was used to measure the biological activity of immunotoxin. SDS-PAGE showed the immunotoxin, free McAb and TCS were in the mixture of conjugation, and the free TCS can be separated by Sephacryl S-200. In Vitro, the lymphocytes of rat can be killed by antiCD4,antiCD8 immunotoxin. The kill capability was relay to the amount of immunotoxin. The authors consider that the immunology unresponsiveness can be induced by antiCD4,antiCD8 immunotoxin. That was useful in induced transplantation tolerance.
ObjectiveTo investigate the role of preoperative peripheral blood CD4/CD8 ratio in predicting the prognosis of patients with coronary atherosclerotic heart disease (CAD) after off-pump coronary artery bypass grafting (OPCABG).MethodsA total of 118 patients with CAD who underwent OPCABG in our hospital from September 2016 to April 2017 were included in the study, including 82 males and 36 females aged 62.74±4.50 years. The primary end point was the incidence of major adverse cardiovascular events (MACE). Patients were divided into a high CD4/CD8 group (≥1.40, 62 patients) and a low CD4/CD8 group (<1.40, 56 patients) according to the results of flow cytometry. The correlation between CD4/CD8 ratio and prognosis of patients after OPCABG and the value of CD4/CD8 ratio for predicting postoperative MACE were evaluated.ResultsMedian duration of follow-up was 23.25 (20.91, 24.70) months, during which 21 patients (17.80%) experienced MACE and 4 patients (3.39%) were lost to follow-up. Kaplan-Meier analysis revealed that high CD4/CD8 group had a significantly higher MACE rate than the low CD4/CD8 group did (log-rank χ2=5.797, P=0.02). The results of adjusted Cox proportional hazards model showed that CD4/CD8 ratio (HR=3.103, 95%CI 1.557-6.187, P<0.01) was an independent risk factor of MACE in patients with CAD after OPCABG. The receiver operating characteristic curve showed that area under curve was 0.778 (95%CI 0.661-0.894, P<0.01), the optimal cut off value was 2.24, the sensitivity was 57.1%, and the specificity was 87.6%.ConclusionPreoperative peripheral blood CD4/CD8 ratio is an independent predictor of MACE after OPCABG in patients with CAD.
ObjectiveTo study the relationship between the expression of sialyl Lewisx (SLeX) antigen and CD44v6 products and biological behaviors in cholangiocarcinomas. MethodsThe expression of SLeX and CD44v6 in 43 cases of cholangiocarcinoma tissue was respectively investigated by catalyzed signal amplification immunohistochemical technique.The relationship between expression of SLeX and CD44v6 and the clinicopathological factors of cholangiocarcinoma was analyzed.ResultsThe positive expression rate of SLeX and CD44v6 in cholangiocarcinoma was 67.4% and 62.8% respectively,which was significantly higher than that in control group (20.0%,P＜0.05).The high level expression of SLeX and CD44v6 were correlated with the TNM phase, differentiation degree,metastasis to lymph nodes and viscera in cholangiocarcinoma (P＜0.05). Moreover,there was a positive correlation between the SLeX and CD44v6 expression in cholangiocarcinoma (r=0.49,P＜0.001).Conclusion Expression of SLeX and CD44v6 could be helpful in predicting the biological behavior and prognosis of cholangiocarcinoma.
Objective To explore the role of CD4+CD25+ Treg cells in chronic pulmonary infection caused by Pseudomonas aeruginosa(PA).Methods Sixty SD rats were randomly divided into a PA group and a control group(n=30 in each group).Chronic lung infection model was established by implantation of silicone tube precoated with PA into the main bronchus.Twenty-eight days later Treg cells in peripheral blood were measured by fluorescence-activated cell sorting（FACS）.Levels of IL-10 and TGF-β in serum were assayed by ELISA.The expression of Foxp3 mRNA in spleen was measured by RT-PCR.Pathological changes of lung tissue were studed by HE staining.Results Treg/CD4+ T cells in the PA group were significantly more than those in the control group[（19.79±6.45）% vs （5.15±0.47）%，Plt;0.05].The levels of IL-10 and TGF-β were (231.52±54.48)pg/mL and （121.05±7.98）pg/mL in the PA group respectively，which were significantly higher than those in the control group[（35.43±23.56）pg/mL and （36.02±8.94）pg/mL].The expression of Foxp3 mRNA in the PA group was significantly higher compared with the control group(0.80±0.044 vs 0.25±0.054，Plt;0.05）.HE staining revealed that PA caused a intensive inflammatory reaction with lymphocytes infiltration.Conclusion CD4+CD25+ Treg cell is up-regulated and plays an important role in chronic lung infection caused by Pseudomonas aeruginosa.
The adhesion of cancer cells to the vascular endothelium during hematogenous metastasis is a crucial first step, involving the interaction of multiple adhesion molecules between cancer cells and endothelial cells. Here, a parallel-plate flow chamber combined with fluorescence microscopy was used to observe the adhesion behavior and subsequent calcium response of MDA-MB-231 cells on different functionalized substrates under flows, revealing the underlying force-regulation mechanism by analyzing and extracting relevant characteristic parameters. Our results demonstrated that fluid shear stress positively regulated the rolling velocity of cells by affecting the dissociation rate constant of CD44/E-selectin, and rapidly activated integrin α5β1 at the sub-second level, slowing down the rolling velocity of cells, but not enough to firm adhesion. Force triggered the calcium response of MDA-MB-231 cells on E-selectin. Furthermore, the activated integrin α5β1 binding with fibronectin enhanced and quickened cellular calcium response with higher activation ratio and peak intensity, and shorter delay time. This study can deepen the understanding of the hematogenous metastasis process of breast cancer cells, and provide reference for relevant clinical treatment strategies and drug development.
ObjectiveTo investigate the expressions of CD133 and CD44 protein in primary lesions of gastric cancer and its clinical significance. MethodsThe expressions of CD44 and CD133 protein in gastric cancer tissues of 100 patients with gastric cancer were detected by immunohistocheimcal stainings.The relation between the expressions of CD44 and CD133 protein and the clinicopathologic characters were analyzed. ResultsBoth CD44 and CD133 protein were expressed on the cell membranes.No correlation were found between CD44/CD133 and the clinicopathologic parameters include gender and age (P > 0.05), but the positive expression rate of CD44/CD133 with diameter>5 cm was significantly higher than that tumor with diameter≤5 cm (CD44 P=0.150;CD133 P=0.056), and correlated with the tissue differentiation (CD44 P=0.008;CD133 P=0.007), vascular invasion (CD44 P=0.043;CD133 P=0.023), lymphatic vessel invasion (CD44 P=0.020;CD133 P=0.044), lymph nodes metastasis (CD44 P=0.002;CD133 P=0.004), inva-sion depth of tumor (CD44 P=0.006;CD133 P=0.021), and pTNM stage (CD44 P=0.034;CD133 P=0.001).No correlation were found between the co-expression of CD44 and CD133 protein and the clinicopathologic parameters include gender, age, tissue differentiation, and vascular invasion (P > 0.05), but the positive co-expression rate of CD44 and CD133 with diameter>5 cm was significantly higher than that tumor with diameter≤5 cm (P=0.010), and correlated with lymphatic vessel invasion (P=0.003), lymph nodes metastasis (P=0.045), invasion depth of tumor (P=0.041), and pTNM stage (P=0.049).The Spearman rank correlation analysis showed that there was positive correlation between the expressions of CD44 and CD133 protein (r=0.207, P=0.039).Univariate analysis showed that lymph nodes metastasis (P < 0.001), pTNM stages (P=0.013), CD44 protein expression (P=0.005), CD133 protein expression (P=0.002), and co-expression of CD44 and CD133 protein (P < 0.001) were significantly correlated with 3-year survival rate of pati-ents with gastric cancer respectively.Logistic regression analysis revealed that lymph node metastasis (P=0.038) was independent risk factor for co-expression of CD44 and CD133 protein.Multivariate analysis with the Cox regression models showed that co-expression of CD44 and CD133 protein (P=0.003) and lymph node metastasis (P=0.006) were significantly associated with poor prognosis. ConclusionsCD44 and CD133 protein may be considered as robust cancer stem cell markers in gastric cancer.The co-expression of CD44 and CD133 protein is the independent prognosis factor for gastric cancer and strongly associated with poor prognosis when they are expressed more high.
Objective To observe the therapeutic effect of mensenchymal stem cells (MSCs) for experimental autoimmune uveitis (EAU). Methods MSCs were obtained from Wistar rats and selected by plastic adherence. Lewis rats were divided into treatment group and control group, six rats in each group. EAU models were induced by immunization with an emulsion (0.2 ml) containing 30 mu;g interphotoreceptor retinoid-binding protein derived peptide R16 and complete Freundprime;s adjuvant. The clinical manifestations of two groups were observed. Nine to 11 day after modeling, 1 ml MSCs suspension, which contained 5times;106 MSCs, were injected into the rats in treatment group via tail vein, and the rats in control group were given equal volume of phosphate buffer solution. Fifteen day after modeling, the eyes were collected to test the proportion of interferon gamma;, interleukin-17 and Foxp3 positive cells by flow cytometry. The clinical scores were analyzed by mixed linear model and statistical analysis of variance of repeated measurement data. The results of flow cytometry were analyzed using independent-sample t test. Results Six days after immunization, mild dilatation and congestion of iris vascular was observed. Nine days after immunization, mild muddy anterior chamber, myosis and absent pupillary reaction to light were observed. Twelve days after immunization, muddy anterior chamber, occlusion of pupil and dimmed or disappeared red reflex were observed, and then inflammation was slowly reduced. From 11 to 15 days after immunization, the clinical score of treatment group was lower than that in control group, the difference was statistically significant (t=2.42, 2.21, 4.16, 5.24, 4.03; P＜0.05). The results of flow cytometry showed that MSCs treatment could decrease the proportion of CD4＋T cells, Th1 cells and Th17 cells, increase the proportion of Treg cells. Conclusion MSCs treatment can ameliorate EAU, up-regulate the expression of Treg cells and down-regulate the expression of CD4＋T cells, Th1 cells and Th17 cells.