Pretreatment levels of circulating endothelial cells on efficacy of first-line therapy in patients with advanced non-small cell lung cancer

Background:Elevated levels of circulating endothelial cells might reflect significant vascular damage and dysfunction.Recent studies have shown that circulating endothelial cells levels are related to therapeutic responses of tumors,and thus,could be used as an indicator to predict the efficacy of tumor treatments.The purpose of this study was to investigate the correlation and impact of endothelial cells with and on the efficacy of first-line therapy(platinum-based or tyrosine kinase inhibitor drugs treatments)for advanced non-small-cell lung cancer.Methods:We analyzed 45 inpatients who met the inclusion criteria of diagnosis with inoperable non-small-cell lung cancer stages III and IV,in the People’s Hospital of Guangxi Zhuang Autonomous Region from January 2019 to January 2020.The flow cytometry technique was adopted to detect pretreatment levels of circulating endothelial cells in peripheral blood of patients with advanced non-small-cell lung cancer.The pretreatment peripheral blood was collected to analyze the relations of circulating endothelial cells with different clinical characteristics and efficacy.Results:The level of pretreatment circulating endothelial cells was significantly correlated with the efficacy of treatment(P<0.05)but irrelevant to the patient’s physical conditions,pathological type,tumor stage,and pretreatment serum carcinoembryonic antigen(P>0.05).The comparison between the groups of response(complete response+partial response)and nonresponse(stable disease+progressive disease)showed a significant difference in circulating endothelial cells count.Compared with low levels of circulating endothelial cells,a high level of circulating endothelial cells led to a poor efficacy(P<0.05).Conclusion:The level of pretreatment circulating endothelial cells significantly correlated with the efficiency of first-line therapy for non-small-cell lung cancer.Compared with low level of circulating endothelial cells,high level of circulating endothelial cells lead to poor efficacy.Therefore,circulating endothelial cell is indeed an effective indicator for predicting the efficacy of first-line therapy for advanced non-small-cell lung cancer.

Changes of activated circulating endothelial cells and survivin in patients with non-small cell lung cancer after antiangiogenesis therapy

Background Although antiangiogenesis therapy plays an important role in anti-neoplastic treatment with its recognized efficacy and slight adverse effect, there is no prospective clinical trial to define ideal markers for predicting efficacy of antiangiogenic therapy. This study was undertaken to investigate the changes of activated circulating endothelial cells （aCECs） and survivin after anti-angiogenesis therapy and their significance in predicting the efficacy of the therapy. Methods Patients of non-small cell lung cancer （NSCLC） treated with chemotherapy with or without Endostar were observed. The amount of activated CECs was detected by flow cytometry, and the expression of survivin mRNA was determined by real-time polymerase chain reaction （PCR）. Results After treatment, the amount of activated CECs decreased significantly in clinical benefit cases （P=0.021 in chemotherapy alone, P=0.001 in chemotherapy plus Endostar）, increased in disease progressive cases （P=0.015 in chemotherapy alone, but P=0.293 in chemotherapy with Endotatar）. After therapy, the expression of survivin mRNA decreased in clinical benefit cases （P=0.001） and increased in disease progressive cases （P=0.018）. A positive correlation was found between activated CECs and survivin in the chemotherapy group pre- and post-therapy （P=0.001 and 0.021, respectively）, but only in the chemotherapy with Endostar group pre-therapy （P=0.030） rather than post-therapy. A positive correlation was found between the decreased activated CECs after therapy and time to progression （TTP） （r=0.322, P=0.012）; a negative correlation was found between the amount of survivin mRNA in serum post-therapy and -l-I-P（r= -0.291, P=0.048）. Conclusions Activated CECs and survivin may be ideal markers forecasting efficacy and prognosis of NSCLC. The former can reflect more sensitively antiangiogenic efficacy and the latter is more sensitive to shrinkage or swelling of tumors. Their combination can evaluate more accurately the efficacy of antiangiogenic therapy of NSCLC.

Correlation between Increased Circulating Endothelial Progenitor Cells and Stage of non-Hodgkin Lymphoma

This study aims to examine the levels of circulating endothelial progenitor cells （cEPCs） in the peripheral blood of patients with non-Hodgkin lymphoma （NHL） and their correlation with the tumor stage. Forty-one patients with biopsy-proven NHL and 16 healthy individuals were recruited. Pe- ripheral blood mononuclear cells （PBMCs） were isolated by density gradient centrifugation, and cEPCs were characterized by triple staining using antibodies against CD133, CD34 and vascular endothelial growth factor receptor-2 （VEGFR-2, CD309） and quantified by flow cytometry. In NHL patients, the number of cEPCs was significantly greater than in control group （P=-0.000）. The cEPCs counts in patients with NHL of stage III-1V were significantly greater than in stage I -II （P=-0.010）. FACS analysis revealed that the number of cEPCs in NHL patients had no correlation with the gender （P=0.401） or the pathological category （P=0.852）. It was suggested that the over-expression of cEPCs in NHL patients may serve as a novel biomarker for disease progression in NHL.

Effect of Composite Salviae Dropping Pill on Endothelin Gene Expression in Circulating Endothelial Cells of Patients with Coronary Heart Disease

Objective: To explore the effect of composite Salviae droping pill(CSDP) on endothelin (ET-1) gene expression in circulating endothelial cells. Mothods: Seventy cases of stable angina pectoris were randomly divided into two groups, the CSDP group and the isosorbide dinitrate (ID) group. They were treated with CSDP and ID respectively,their ET-1 gene expression in endothelial cells of peripheral circulation was measured before and after treatment by reverse transcriptase-polymerase chain reaction(RT-PCR) and compared between the two groups as well as with that of healthy subjects. Results: Electrophoresis banding of 546 bp cDNA procured from 59 cases of 70 patients was positive, while no positive banding was obtained from the healthy subjects. Six cases from the 29 patients treated with CSDP had their banding turned to negative,while in the ID group,no one turned to negative after treatment. And ET-1 PCR product (absorbed optic density) in the CSDP group was markedly lower than that in the ID group, P＜0.05. Conclusion: DSP could directly inhibit ET-1 gene expression in endothelial cells of peripheral circulation.

Endothelial progenitor cells mobilization after maximal exercise according to heart failure severity

BACKGROUND Vascular endothelial dysfunction is an underlying pathophysiological feature of chronic heart failure(CHF).Patients with CHF are characterized by impaired vasodilation and inflammation of the vascular endothelium.They also have low levels of endothelial progenitor cells(EPCs).EPCs are bone marrow derived cells involved in endothelium regeneration,homeostasis,and neovascularization.Exercise has been shown to improve vasodilation and stimulate the mobilization of EPCs in healthy people and patients with cardiovascular comorbidities.However,the effects of exercise on EPCs in different stages of CHF remain under investigation.AIM To evaluate the effect of a symptom-limited maximal cardiopulmonary exercise testing(CPET)on EPCs in CHF patients of different severity.METHODS Forty-nine consecutive patients(41 males)with stable CHF[mean age(years):56±10,ejection fraction(EF,%):32±8,peak oxygen uptake(VO2,mL/kg/min):18.1±4.4]underwent a CPET on a cycle ergometer.Venous blood was sampled before and after CPET.Five circulating endothelial populations were quantified by flow cytometry:Three subgroups of EPCs[CD34+/CD45-/CD133+,CD34+/CD45-/CD133+/VEGFR2 and CD34+/CD133+/vascular endothelial growth factor receptor 2(VEGFR2)]and two subgroups of circulating endothelial cells(CD34+/CD45-/CD133-and CD34+/CD45-/CD133-/VEGFR2).Patients were divided in two groups of severity according to the median value of peak VO2(18.0 mL/kg/min),predicted peak VO2(65.5%),ventilation/carbon dioxide output slope(32.5)and EF(reduced and mid-ranged EF).EPCs values are expressed as median(25th-75th percentiles)in cells/106 enucleated cells.RESULTS Patients with lower peak VO2 increased the mobilization of CD34+/CD45-/CD133+[pre CPET:60(25-76)vs post CPET:90(70-103)cells/106 enucleated cells,P<0.001],CD34+/CD45-/CD133+/VEGFR2[pre CPET:1(1-4)vs post CPET:5(3-8)cells/106 enucleated cells,P<0.001],CD34+/CD45-/CD133-[pre CPET:186(141-361)vs post CPET:488(247-658)cells/106 enucleated cells,P<0.001]and CD34+/CD45-/CD133-/VEGFR2[pre CPET:2(1-2)vs post CPET:3(2-5)cells/106 enucleated cells,P<0.001],while patients with higher VO2 increased the mobilization of CD34+/CD45-/CD133+[pre CPET:42(19-73)vs post CPET:90(39-118)cells/106 enucleated cells,P<0.001],CD34+/CD45-/CD133+/VEGFR2[pre CPET:2(1-3)vs post CPET:6(3-9)cells/106 enucleated cells,P<0.001],CD34+/CD133+/VEGFR2[pre CPET:10(7-18)vs post CPET:14(10-19)cells/106 enucleated cells,P<0.01],CD34+/CD45-/CD133-[pre CPET:218(158-247)vs post CPET:311(254-569)cells/106 enucleated cells,P<0.001]and CD34+/CD45-/CD133-/VEGFR2[pre CPET:1(1-2)vs post CPET:4(2-6)cells/106 enucleated cells,P<0.001].A similar increase in the mobilization of at least four out of five cellular populations was observed after maximal exercise within each severity group regarding predicted peak,ventilation/carbon dioxide output slope and EF as well(P<0.05).However,there were no statistically significant differences in the mobilization of endothelial cellular populations between severity groups in each comparison(P>0.05).CONCLUSION Our study has shown an increased EPCs and circulating endothelial cells mobilization after maximal exercise in CHF patients,but this increase was not associated with syndrome severity.Further investigation,however,is needed.

Correlation between circulating endothelial cell level and acute respiratory distress syndrome in postoperative patients

BACKGROUND Acute respiratory distress syndrome(ARDS)is injury of alveolar epithelial cells and capillary endothelial cells caused by various factors,including endogenous and exogenous lung factors,leading to diffuse pulmonary interstitial and alveolar edema,and acute respiratory failure.ARDS involves alveolar epithelial cells and pulmonary interstitial capillary endothelial cells.Circulating endothelial cells(CECs)are the only marker that directly reflects vascular endothelial injury in vivo.There have been few studies on the correlation between peripheral blood CECs and ARDS at home and abroad.The lungs are the organs with the highest capillary density and the most endothelial cells,thus,it is speculated that when ARDS occurs,CECs are stimulated and damaged,and released into the circulatory system.AIM To explore the correlation between CEC level and severity of ARDS in patients postoperatively.METHODS Blood samples were collected from all patients on day 2(d2)and day 5(d5)after surgery.The control group comprised 32 healthy volunteers.Number of CECs was measured by flow cytometry,and operation time was recorded.Changes in various indexes of patients were monitored,and diagnosis of ARDS was determined based on ARDS Berlin definition.We comprised d2 CECs in different groups,correlation between operation time and d2 CECs,ARDS of different severity by d2 CECs,and predictive value of d2 CECs for ARDS in postoperative patients.RESULTS The number of d2 CECs in the ARDS group was significantly higher than that in the healthy control group(P<0.001).The number of d2 CECs in the ARDS group was significantly higher than that in the non-ARDS group(P<0.001).The number of d2 CECs in the non-ARDS group was significantly higher than that in the healthy control group(P<0.001).Operation time was positively correlated with number of CECs on d2(rs=0.302,P=0.001).The number of d2 CECs in the deceased group was significantly higher than that in the improved group(P<0.001).There was no significant difference in number of d2 CECs between patients with mild and moderate ARDS.The number of d2 CECs in patients with severe ARDS was significantly higher than that in patients with mild and moderate ARDS(P=0.041,P=0.037).There was no significant difference in number of d5 and d2 CECs in the non-ARDS group after admission to intensive care.The number of d5 CECs was higher than the number of d2 CECs in the ARDS improved group(P<0.001).The number of d5 CECs was higher than the number of d2 CECs in the ARDS deceased group(P=0.002).If the number of CECs was>1351/mL,sensitivity and specificity of predicting ARDS were 80.8%and 78.1%,respectively.CONCLUSION Changes in number of CECs might predict occurrence and adverse outcome of ARDS after surgery,and higher numbers of CECs indicate worse prognosis of ARDS.

Decreased and dysfunctional circulating endothelial progenitor cells in patients with chronic obstructive pulmonary disease

Background It has been widely demonstrated that endothelial progenitor cells are involved in several diseases and that they have therapeutic implications. In order to define the altered pulmonary vascular homeostasis in chronic obstructive pulmonary disease, we sought to observe the level and functions of circulating endothelial progenitor calls in patients with chronic obstructive pulmonary disease. Methods The total study population included 20 patients with chronic obstructive pulmonary disease and 20 control subjects. The number of circulating endothelial progenitor cells （CD34＋/CD133＋/IVEGFR-2＋cells） was counted by flow cytometry. Circulating endothelial progenitor cells were also cultured in vitro and characterized by uptake of Dil- acLDL, combining with UEA-I, and expression of von Willebrand factor and endothelial nitric oxide synthase. Adhesion, proliferation, production of nitric oxide, and expression of endothelial nitric oxide synthase and phosphorylated-endothelial nitric oxide synthase were detected to determine functions of circulating endothelial progenitor cells in patients with chronic obstructive pulmonary disease. Results The number of circulating endothelial progenitor cells in the chronic obstructive pulmonary disease group was lower than in the control group： （0.54±0.16）% vs. （1.15±0.57）%, P 〈0.05. About 80% of adherent peripheral blood mononuclear cells cultured in vitro were double labeled with Dil-acLDL and UEA-I. The 92% and 91% of them were positive for von Willebrand factor and endothelial nitric oxide synthase, respectively. Compared with the control, there were significantly fewer adhering endothelial progenitor cells in chronic obstructive pulmonary disease patients： 18.7±4.8/field vs. 45.0±5.9/field, P 〈0.05. The proliferation assay showed that the proliferative capacity of circulating endothelial progenitor cells from chronic obstructive pulmonary disease patients was significantly impaired： 0.135±0.038 vs. 0.224±0.042, P 〈0.05. Furthermore, nitric oxide synthase （112.06±10.00 vs. 135.41±5.38, P 〈0.05）, phosphorylated endothelial nitric oxide synthase protein expression （88.89±4.98 vs. 117.98±16.49, P 〈0.05） and nitric oxide production （（25.11±5.27） Iμmol/L vs. （37.72±7.10） μmol/L, P 〈0.05） were remarkably lower in endothelial cells from the chronic obstructive pulmonary disease group than the control. Conclusion Circulating endothelial progenitor cells were decreased and functionally impaired in patients with chronic obstructive pulmonary disease.

Precise capture of circulating endometrial cells in endometriosis

Background:Endometriosis(EM)is a complex benign gynecological disease,but it has malignant biological behavior and can invade any part of the body.Clinical manifestations include pelvic pain,dysmenorrhea,infertility,pelvic nodules,and masses.Our previous study successfully detected circulating endometrial cells(CECs)in the peripheral blood of patients with EM.The purpose of this study is to overcome the limitation of cell size in the previous microfluidic chip method,to further accurately capture CECs,understand the characteristics of these cells,and explore the relationship between CECs and the clinical course characteristics of patients with EM.Methods:Human peripheral venous blood used to detect CECs and circulating vascular endothelial cells(CVECs)was taken from EM patients(n=34)hospitalized in the Peking University People’s Hospital.We used the subtraction enrichment and immunostaining fluorescence in situ hybridization(SE-iFISH)method to exclude the interference of red blood cells,white blood cells,and CVECs,so as to accurately capture the CECs in the peripheral blood of patients with EM.Then we clarified the size and ploidy number of chromosome 8 of CECs,and a second grouping of patients was performed based on clinical characteristics to determine the relationship between CECs and clinical course characteristics.Results:The peripheral blood of 34 EM patients and 12 non-EM patients was evaluated by SE-iFISH.Overall,34 eligible EM patients were enrolled.The results showed that the detection rates of CECs were 58.8%in EM patients and 16.7%in the control group.However,after classification according to clinical characteristics,more CECs could be detected in the peripheral blood of patients with rapidly progressive EM,with a detection rate of 94.4%(17/18).In total,63.5%(40/63)of these cells were small cells with diameters below 5μm,and 44.4%(28/63)were aneuploid cells.No significant association was found between the number of CECs and EM stage.Conclusion:The number and characteristics of CECs are related to the clinical course characteristics of patients with EM,such as pain and changes in lesion size,and may be used as biomarkers for personalized treatment and management of EM in the future.

Assessment of Reversibility in Pulmonary Hypertension Related to Congenital Heart Disease by Using Biomarkers and Clinical Features

Background:Reversibility of pulmonary hypertension(PH)is closely related to the treatment options for and prognosis of children with congenital heart disease.Objective:We combined patient-specific clinical features including diagnosis,age and echocardiographic results,and biomarkers of pulmonary vascular dysfunction to explore the noninvasive methods that can be used to accurately evaluate the reversibility of pulmonary hypertension in congenital heart disease(PH-CHD).Methods:Based on the preoperative systolic pulmonary arterial pressure(sPAP),70 CHD patients were divided into normal,PH-CHD suspected,and confirmed groups.Additionally,biomarkers of circulating endothelial cells(CECs),endothelin-1(ET-1),and endothelial nitric oxide synthase(eNOS)were detected.Patients were categorized into reversible(RPH)and irreversible(IRPH)groups according to the sPAP 6 months after surgery.Risk stratification was performed according to the clinical features and biomarkers.Results:CECs and ET-1 levels in the confirmed group were significantly higher.eNOS was higher in the confirmed and suspected groups than that in the normal group.CECs in the IRPH group were significantly higher compared to the RPH group.No such intergroup differences were observed with respect to ET-1 and eNOS levels.The ROC curve showed that the risk stratification was of high diagnostic value to evaluate reversibility.Conclusion:The CECs,eNOS,and ET-1 were closely related with PH-CHD.CECs and risk stratification have high practical value in assessing the reversibility of PH-CHD.

Protecdive Effects of Radix Astrsgalus on Vascular Endotbelial Cells of Patients with Binswanger's Disease

Objective: To compare the effects of Radix Astragalus (RA) on vascular endothelial cells inBinswanger's disease (BD) patients with Radix Salviae miltiorrhizae (RSM). Methods: There were 37 patientswith BD in the treated group and 37 healthy subjects in the control group. Thirty-seven patients were furtherrandomly subdivided into two groups: RA group (19 patients) and RSM group (18 patients). Circulating endothelial cells (CEC) and the levels of endothelin--1 (ET--1 ), nitric oxide (NO) and malondialdehyde (MDA) inthe blood of internal jugular vein which were examined before and after treatments. Results: When comparedwith those of the control group, CEC counts, ET-1 and MDA levels in plasma increased significantly, meanwhile serum NO concentration decreased significantly in the treated group. When compared with those of pretreatment, CEC counts, ET-1 and MDA decreased significantly and serum NO concentration increased significantly after treatment in RA groUp. There were no significant changes of these indices in RSM group after treatment. Conclusions: There are damage and dysfunction of vascular endothelial cells in patients with BD. RA injection is an effective drug to protect vascular endothelial cells of BD patients.