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AIM: To evaluate the expression of fibroblast growth factor receptor 3 (FGFR3) in acute lymphocytic leukemia (ALL) patients and its contribution to the proliferation of circulating endothelial cells (CECs).METHODS: The mRNA expression levels of FGFR3 in 44 patients with ALL were assayed by RT-PCR.Overall survival (OS) rates of the patients in FGFR3+ group and FGFR3-group were estimated by Kaplan-Meier analysis.The CECs were sorted from peripheral blood by magnetic-activated cell sorting and then counted by 3-color flow cytometry.The cell counts, antigen expression, growth curve and colony forming rate of the CECs in the 2 groups were determined.The FGFR3 expression of CECs was identified by immunofluorescence staining.RESULTS: The positive rate of FGFR3 mRNA expression was 43.2% in 44 ALL patients with normal karyotype.T-ALL expressed higher level of FGFR3 than B-ALL (P<0.05).FGFR3 was over-expressed in ALL patients with bone marrow blast proportion ≥5% (P<0.05).The probability of OS was significantly lower in FGFR3+ group than that in FGFR3-group (P<0.05).The sorted CECs highly expressed CD31, CD144, VEGFR-2 and CD146, and rarely expressed CD45.The counts of CECs and expression level of CD133 significantly increased in FGFR3+ group compared with FGFR3-group.The same result of the amount of colony formation was observed (P<0.05).There was significant difference at 3 time points of cultured CECs count in vitro between FGFR3+ group and FGFR3-group (P<0.05).The positive rate of FGFR3 expression of CECs from 19 ALL-FGFR3+ patients was (29.00±15.71)%.CONCLUSION: The over-expression of FGFR3 gene in ALL may be helpful to evaluate the prolife-ration of CECs, and become a double target with anti-tumor and anti-angiogenesis effects to offer more choice for molecular therapy in the future.
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AIM:To determine the biological feature of circulating endothelial cells (CECs) in acute promye-locytic leukemia ( APL) patients before and after treatment , and to analyze the relationship between CECs and the clinical characteristics .METHODS: The CECs were sorted from peripheral blood by magnetic-activated cell sorting and then counted by 3-color flow cytometry.The cells were identified by immunofluorescence staining for the expression of CD 146, CD31, CD144, VEGFR-2, CD45 and CD133.The CECs were cultured in vitro, and the tube formation and colony-forming rate were determined .RESULTS:Increased quantity of CECs was observed in CD 34 positive group and group with WBC >10 ×109/L (P<0.05).The quantity of CECs had a significant difference among low risk , medium risk and high risk groups (P<0.05).The positive rate of CD133 and quantity of CECs significantly reduced in 32 APL patients when they gain complete remission after treatment (P<0.05).The amount of tube formation and colony-forming rate were significant-ly reduced after treatment (P<0.05).The ratio of CECs quantity from APL patients after treatment to that before treatment had a negative correlation with arsenic concentration in urine on day 7 during As2O3 treatment (P<0.05).CONCLU-SION:Accurately counting CECs may be helpful for evaluating prognosis and designing treatment strategy .
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We measured circulating endothelial precursor cells (EPCs), activated circulating endothelial cells (aCECs), and mature circulating endothelial cells (mCECs) using four-color multiparametric flow cytometry in the peripheral blood of 84 chronic myeloid leukemia (CML) patients and 65 healthy controls; and vascular endothelial growth factor (VEGF) by quantitative real-time PCR in 50 CML patients and 32 healthy controls. Because of an increase in mCECs, the median percentage of CECs in CML blast crisis (0.0146%) was significantly higher than in healthy subjects (0.0059%, P<0.01) and in the accelerated phase (0.0059%, P=0.01). There were no significant differences in the percentages of CECs in chronic- or active-phase patients and healthy subjects (P>0.05). In addition, VEGF gene expression was significantly higher in all phases of CML: 0.245 in blast crisis, 0.320 in the active phase, and 0.330 in chronic phase patients than it was in healthy subjects (0.145). In conclusion, CML in blast crisis had increased levels of CECs and VEGF gene expression, which may serve as markers of disease progression and may become targets for the management of CML.
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Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Blast Crisis/pathology , Endothelial Cells/pathology , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/pathology , Neoplastic Cells, Circulating/pathology , Vascular Endothelial Growth Factor A/genetics , Biomarkers, Tumor/analysis , Blast Crisis/blood , Blast Crisis/genetics , Case-Control Studies , Cell Count , Flow Cytometry/methods , Gene Expression/genetics , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/blood , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/genetics , Neovascularization, Pathologic/pathology , Real-Time Polymerase Chain Reaction , Reference Values , Statistics, Nonparametric , Vascular Endothelial Growth Factor A/analysisABSTRACT
Objective To compare the count of circulating endothelial cells (CECs) between patients with reversible pulmonary hypertension (RPH) and irreversible pulmonary hypertension (iRPH) by flow cytometry in order to find a new biomarker to distinguish RPH from iRPH.Methods Ninety pulmonary hypertension associated with congenital heart disease patients treated in our hospital between September 1,2013 to March 31,2014 were enrolled in this study.According to the mean pulmonary arterial pressure (mPAP) measured by echocardiography in six months after treatment,the patients were separated into 2 groups:iRPH group(mPAP≥50 mmHg),RPH group(mPAP ＜50 mmHg).Results Totally the clinical data of 87 patients were enrolled in the statistic analysis,86 patients finished the follow-up in 6 months after the surgery,1 patient died from pulmonary hypertensive crisis.The count of CECs was not correlated with the extent of pulmonary hypertension(P =0.925).The number of CECs was higher in iRPH group than that of RPH group(0.46‰ vs.0.09‰,Z =-5.021,P =0.000).And also the age of patients in iRPH group was elder than that of patients in RPH group [(43.1 ± 37.4) months vs.(9.3 ± 12.6) months,t =-5.079,P =0.000].Conclusion The count of CECs is significantly increased in iRPH patients.It could be used as one biomarker to distinguish RPH from iRPH.
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Objective To explore changes in circulating endothelial cells(CECs)numbers in neonatal hypoxemia and its clinical significance.Methods Hladovec method was used to measure the quantity of CECs in 40 cases of neonatus with different degree of hypoxemia(hypoxemia group),and 20 matched healthy newborns were enrolled as control group.Results The numbers of CECs in hypoxemia group were much more than those in control group,and the difference was statistically significant.Meanwhile,there were significant differences in CECs numbers between moderate group and mild group(t =13.43,P <0.05),as well as moderate group and severe group(t=7.698,P <0.05).Conclusion The quantity of circulating ECEs in hypoxic neonatus is correlated with the hy-poxic severity,which may be used as early diagnostic indicator for hypoxemia,and could provide new scientific evidences for early diagnosis,illness judgment and curative effect evaluation.
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Objective: This study aimed to examine the number of activated circulating endothelial cells (aCECs) in the peripheral blood of patients with non-small cell lung cancer (NSCLC), and investigate the relationship among aCECs, anti-angiogenic therapy, and prognosis of NSCLC patients. This study also aimed to identify novel predictive markers for anti-angiogenic therapy, and provide basic data and experimental basis for establishing an evaluation system for this therapy. Methods: A total of 142 NSCLC patients were randomly divided into the chemotherapy group (Group 1) and combined therapy group (i.e., chemotherapy plus endostatin, Group 2). The number of aCECs was measured using flow cytometry by detecting the expression status of CD105 and CD146 in the peripheral blood. The correlation between the changes in aCECs and efficacy of drug treatment was statistically analyzed using SPSS software. Results:The number of aCECs in Group 2 increased significantly at 8 and 29 d, two cycles, 50 and 71 d, and four cycles after treatment, respectively (P<0.05). In particular, aCECs amount in cases of progressive disease increased more significantly after combined therapy (P<0.05). A negative correlation was found between the treatment cycle and difference in aCECs amount before and after therapy (r=-0.970, P=0.001). A negative correlation was also observed between the difference in aCECs amount and time to tumor progression (TTP) (r=-0.351, P=0.039). Therefore, the difference in aCECs amount before and after therapy could serve as an important predictor for TTP in NSCLC patients. Conclusion:CD105 and CD146 reflected the activation status of endothelial cells, and responded to the drug treatment. Thus, CD105 and CD146 could act as ideal markers for aCECs. The number of aCECs increased during cancer progression, but significantly decreased after long-term treatment. Therefore, the change in aCECs amount may be a useful marker in predicting the efficacy of anti-angiogenic therapy.
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Circulating endothelial cells(CEC)are mature endothelial cells,which have been shed from the vascular cell lining and enter into blood circulation.Rare in healthy individuals,increased CEC in peripheral blood reflects significant vascular damage and dysfunction.Increased CEC have been documented in many human diseases characterized as vascular damage,including different types of tumors.Clinical data suggest that CEC count is a promising tool for monitoring disease activity with potential to assess tumor prognosis and response to treatment.
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ObjectiveTo investigate the relationship between the change of circulating endothelial cell (CEC) level and coronary artery lesion (CAL) of Kawasaki disease (KD),and to further explore the method for early diagnosis of KD.MethodsThirty KD children were recruited for study,including 23 children with complete type of KD and seven children with incomplete KD.According to the results of echocardiography,the KD group was divided into CAL group (9 cases) and non-coronary artery lesion (NCAL)group (21 cases).Ten healthy children were enrolled as control group.Double-blind and controlled trial was conducted,and Hladovec method was applied for CEC counting.Results The CEC level was ( 1.09 ±0.60) × 107/L in KD group,which was higher than that of control group [ (0.38 ±0.14) × 107/L],and the difference was statistically significant ( t =2.85,P ＜ 0.01 ).The CEC level in the CAL group [ ( 1.84 ± 0.24) × 107/L] was higher than that of the NCAL group[ (2.01 ±0.38) × 107/L],and the difference was statistically significant ( t =2.24,P ＜ 0.05 ).The CEC level was ( 1.16 ± 0.63 ) × 107/L in the complete type of KD group and (0.83 ± 0.45 ) × 107/L in the incomplete KD group,which showed no significant difference between the two groups ( t =1.86,P ＞ 0.05 ).CondusionCEC level was elevated significantly in the acute phase of KD.The CEC level in CAL group was higher than that of NCAL group in acute phase.CEC level detection may be helpful for the early diagnosis of KD.
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Objective To evaluate the mid-term and long-term efficacy of recombinant human endostatin (rh-endostatin, EndostarTM) in combination with docetaxel and carboplatin (TP) regimen as a postoperative adjuvant treatment for non-small-cell lung cancer (NSCLC). Methods A randomized controlled trial was conducted in 76 patients who were admitted from October 2006 to July 2008. These patients were diagnosed with NSCLC and experienced complete excision. They were randomly divided into two groups (n=38): Endostar™ plus TP regimen was adopted for the treatment group, while only the TP regimen was used for the control group. After 40 months of postoperative followup, the difference in disease-free survival (DFS) and overall survival (OS) between the two groups was evaluated, and the number of circulating endothelial cells (CECs) and tumor microvessel density (MVD) were measured. Results DFS was significantly longer in the treatment group than in the control group (39.4 months versus 27.6 months, P<0.05). The three-year OS rate of the treatment group was significantly higher than that of the control group (89.4% versus 57.9%, P<0.05). The CECs decreased in both groups after the treatment, while the decline in the treatment group was more significant compared with the control group after four cycles of chemotherapy (P<0.05). There was a significant difference in the MVD between stages I, II, and III (P<0.05). The MVD in the patients with positive lymph nodes was significantly higher than that in the patients with negative lymph nodes. The MVD in poor differentiation was higher than that in moderate/good differentiation (P<0.05). Conclusions By comparing with the simple chemotherapy treatment, EndostarTM plus TP regimen as an adjuvant treatment was able to prolong the DFS and improve the three-year OS rate of NSCLC patients. CEC is a good indicator for predicting the efficacy of chemotherapy plus anti-angiogenesis treatment.
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Circulating endothelial cells (CEC) are endothelial cells which are detected in the peripheral blood. There are very few CEC in healthy adults while the number is obviously increasing in patients with arthrosclerosis, diabetes mellitus, lupus erythematosus, et al. Nowadays, flow cytometry analysis and immunomagnetic isolation for CEC are employed successfully in clinic and scientific research. Several research findings have confirmed that there is intimate relation between CEC and tumorigenesis. Because of the important role in angiogenesis and tumor growth, CEC would be a perspective tumor marker in antiangiogenesis and would also predict the chemotherapy efficacy.
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Objective To explore the importance effect of circulating endothelial cells(CEC)in the pathogenesis of Henoch-Schonlein purpura(HSP)by determining the level of CEC in children with HSP,and to explore the relationship between CEC and therapeutic effect of HSP and the relationship between CEC and Henoch-Schonlein purpura nephritis(HSPN).Methods Sixty-six inpatients with HSP and 30 children as healthy control group were selected in the First Affiliated Hospital of Zhengzhou University from May to Dec.2008.The blood of all children,including the children in healthy control group,were sampled 1 mL in the early morning.And the CEC of all children were measured by flow cytometry method.The variation of CEC was analyzed in patients with HSP at acute stage and in healthy control group,in patients with HSP and in patients with HSPN,and in patients with HSP without renal damage at recovery stage and in patients with HSPN at recovery stage.SPSS 13.0 statistical software was used to analyze the data.Results CEC in patients with HSP at acute stage[(47.934 5?16.902 6)%]was significantly higher than that in healthy control group[(37.436 7?10.200 7)%](t=2.900 P0.05).CEC in patients with HSPN at recovery stage[(50.258 8?12.824 2)%]was significantly higher than that in patients with HSP without renal damage at recovery stage [(40.864 0?8.165 2)%](t=2.906 P
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Objective To isolate CESs by dynabeads coated with the specific antibody of CD146 from peripheral blood and its origin was identified.Methods One mL blood with or without admixed HUVECs was diluted(1∶2) in PBS-0.1% BSA.Anti-CD146-coated Dynabeads were added and incubated for 30mins at(4 ℃).Cells bound to anti-CD146-coupled beads were separated from blood in Dynal MPC and then washed and resuspended in(4 mL) buffer.After 4 additional washes with PBS-0.1% BSA using the magnet,the rosetted cells were flushed from the tube wall with (100 ?L) of PBS-BSA with acridine orange or Giemsa,and counted in a hemocytometer in a inverse phase contrast fluorescence microscope.Results The amount of CECs in healthy adult was 10.5(6～16.5)/mL(n=42).The recovery rate was 91%.Isolated CECs were confirmed by positive expression of vWF and CD31.Conclusion Isolation of CECs from blood by immunomagnetic beads coated with antibody of CD146 is characterized by its accuracy,time-saving,high recovery rate,less contaimination of blood and less damage to CECs.It can be used to quantify CECs and to analyze the functional performance of CECs from patients with vascular injury diseases.
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Objective: To examine the effect of Benapril on atherogenesis and the plaque rupture in rabbits.Methods: Thirty four healthy male New Zealand white rabbits were randomly divided into 4 groups.Group C (control group) was fed normal diet for 10 weeks,group HL fed 1% cholesterol diet,group B fed 1% cholesterol diet and Benapril 5 mg/d.Ten rabbits of plaque rupture group were fed 1% cholesterol diet for 10 weeks with atherosclerosis induced by left carotid iliac damage.Ten weeks after the initiation of the diet,an angioplasty was performed.After angioplasty,the surviving rabbits( n =10) were randomized to receive benapril(5 mg/d,each) supplementation in drinking(B group, n =4) or no treatment(untreated group, n =4). The levels of CEC were measured by morphometrical counts at different periods(the 5th week,the 10th week).Before and 5,10 weeks after experiment,fasting blood samples were collected for serum total cholesterol(TC),high density lipoprotein cholesterol(HDL C),low density lipoprotein cholesterol(LDL C),and triglyceride(TG) assay.The levels of plasma circulating endothelial cells(CEC) were measured by morphometrical counts and the levels of plasma von Willebrand factor(vWF) were determined by ELISA.After sacrificing the separated, entire aortas were stained with oil red O and then processed for histological examination;planimetry was done with a computer system.Endothelial cells were confined by the presence of Factor Ⅷ related antigen as the specific cell marker.Results: The serum TC,TG,LDL C increased progressively in group B and group HL,but the levels of vWF in group HL were significantly higher than that in group B on the 5th and 10th respetively( P
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Objective: To analyze the circulating endothelial cells (CECs) and circulating endothelial precursors (CEPs) in peripheral blood of tumor patients. Methods: CECs and CEPs were enumerated in 57 tumor patients and 15 healthy controls by 3-color flow cytometry. The serum level of angiogenesis related cytokines(VEGF, bFGF)was determined by ELISA method. Results: In tumor patients, mean value of CECs and CEPs was (0.378?0.047)% and (0.059?0.013)% respectively. The serum level of VEGF and bFGF were (295.58?59.56) ng/L and (28.73?7.40) ng/L. The amount of CECs/CEPs positively correlated with the serum level of VEGF and bFGF. Conclusions: The CECs/CEPs and serum level of VEGF and bFGF of tumor patients was higher than those of normal controls. VEGF and bFGF may participate into the course of endothelial progenitors mobilization.
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Objectives To search for the pathogenesis of vaseular endothelial injury and its clinical significance.Methods 36 patients with acute lower respiratory trect infection(ALRI)and 30 healthy controls were envolved. Circulating endothelial cells (CEC)and nitric oxide (NO) levels were tested.Results Circulating nitrite/nitra levels,the stable metabolic products of NO,were found to be significantly higher in the patients with ALRI (44.6?22.6umol/L) than that in the controls (24.5?14.1umoI /L, P
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Objective:Analyze the influence of hematopoietic stem cell mobilization on circulating endothelial cells (CECs) and circulating endothelial precursors (CEPs).Methods:CECs and CEPs were enumerated in 68 tumor patients and 15 healthy controls by 3-color flow cytometry. 11 cases underwent PBSC (peripheral blood stem cell) mobilization by combination chemotherapy and G-CSF (granulocyte colony-stimulating factor).Results:CECs and CEPs in tumor patients were 0.378%?0.103% and 0.059%?0.013% respectively,which were higher than that of normal controls.CECs/CEPs in peripheral blood (PB) were increased after G-CSF mobilization.Conclusion:Hematopoietic and endothelial progenitors can be mobilized into the PB by treatment with growth factor G-CSF.