Circulating endothelial and progenitor cells:Evidence from acute and long-term exercise effects

Circulating bone-marrow-derived cells,named endothelial progenitor cells(EPCs),are capable of maintaining,generating,and replacing terminally differentiated cells within their own specific tissue as a consequence of physiological cell turnover or tissue damage due to injury.Endothelium maintenance and restoration of normal endothelial cell function is guaranteed by a complex physiological procedure in which EPCs play a significant role.Decreased number of peripheral blood EPCs has been associated with endothelial dysfunction and high cardiovascular risk.In this review,we initially report current knowledge with regard to the role of EPCs in healthy subjects and the clinical value of EPCs in different disease populations such as arterial hypertension,obstructive sleep-apnea syndrome,obesity,diabetes mellitus,peripheral arterial disease,coronary artery disease,pulmonary hypertension,and heart failure.Recent studies have introduced the novel concept that physical activity,either performed as a single exercise session or performed as part of an exercise training program,results in a significant increase of circulating EPCs.In the second part of this review we provide preliminary evidence from recent studies investigating the effects of acute and long-term exercise in healthy subjects and athletes as well as in disease populations.

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.

Erythropoietin Receptor Positive Circulating Progenitor Cells and Endothelial Progenitor Cells in Patients with Different Stages of Diabetic Retinopathy

Objective To investigate the possible involvement of erythropoietin （EPO）/erythropoietin receptor （EPOR） system in neovascularization and vascular regeneration in diabetic retinopathy （DR）. Methods EPOR positive circulating progenitor cells （CPCs： CD34^＋） and endothelial progenitor cells （EPCs： CD34^＋KDR^＋） were assessed by flow cytometry in type 2 diabetic patients with different stages of DR. The cohort consisted of age- and sex-matched control patients without diabetes （n=7）, non-prolif- erative DR （NPDR, n=7）, proliferative DR （PDR, n=8）, and PDR complicated with diabetic nephropathy （PDR-DN, n=7）. Results The numbers of EPOR^＋ CPCs and EPOR^＋ EPCs were reduced remarkably in NPDR corn pared with the control group （both P（0.01）, whereas rebounded in PDR and PDR-DN groups in varying degrees. Similar changes were observed in respect of the proportion of EPOR^＋ CPCs in CPCs （NPDR vs. control, P（0.01） and that of EPOR^＋ EPCs in EPCs （NPDR vs. control, P〈0.05）. Conclusion Exogenous EPO, mediated via the EPO/EPOR system of EPCs, may alleviate the impaired vascular regeneration in NPDR, whereas it might aggravate retinal neovascularization in PDR due to a rebound of EPOR^＋ EPCs associated with ischemia.

Effect of Chinese Herbs for Activating Blood Circulation,Removing Stasis and Supplementing Qi on the Circulating Endothelial Cells in Patients with Unstable Angina Pectoris

Objective: To observe the effect and clinical significance of circulating endothelial cells (CEC) in the pathogenesis of coronary heart disease with unstable angina pectoris (CHD-UAP), and to explore the protective effect of Chinese herbs for activating blood circulation, removing stasis and supplementing Qi (CH) on CHD-UAP patient's CEC. Methods: Sixty patients with diagnosis of CHD-UAP confirmed and differentiated to be Qi-deficiency and blood stasis by TCM were randomly divided into two groups and treated, on the basis of Western drug-therapy, with Tongxinluo capsule (通心络胶囊, TXL) and Huoxue Tongmai capsule (活血通脉胶囊, HXTM) respectively by way of oral taking three times a day, 3 capsules every time, with 1 month as one therapeutic course. The number of CEC in patients' blood circulation was counted before and after treatment. Besides, the number of CEC in 30 healthy persons was also counted for control. Results: The number of CEC in CHD-UAP patients was significantly higher than that in the healthy persons (P<0.01). After the patients were treated with CH, either TXL or HXTM, it significantly decreased (P<0.01)with insignificant difference between the two treated groups. Conclusion: CEC in CHD-UAP patients is severely damaged and endothelial function in disorder, Chinese herbs have protective effect on patients' CEC.

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.

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.

卡铂联合多西他赛治疗非小细胞肺癌CECs变化

目的:探索用卡铂联合多西他赛作为一线方案治疗晚期非小细胞肺癌(NSCLC),静脉血中循环内皮细胞(CECs)数目在预测化疗有效性中的作用。方法:采用卡铂联合多西他赛治疗晚期NSCLC病人53例,用流式细胞术测定NSCLC病人外周静脉血中CECs数目,同样方法测定健康人静脉血中CECs数目。结果:NSCLC病人化疗前的外周静脉血中CECs数目[(1 798±970)个/mL]明显高于健康人[(135±150)个/mL]。将化疗前CECs数目视为基线值,Ⅳ期病人的基线CECs数目高于IIIB病人(P=0.036)。基线CECs数目和肿瘤体积呈明显正相关性(rs=0.556,P<0.001)。化疗后病人CECs数目明显下降[化疗第22天为(704±603)个/mL],其中临床获益病人CECs数目下降较明显,病情进展(PD)病人CECs数目无明显下降。临床获益病人基线CECs数目明显高于PD病人(P=0.011)。结论:CECs数目可用来预测卡铂联合多西他赛治疗晚期NSCLC的有效性。

子痫前期患者血浆可溶性内皮细胞蛋白C受体、外周血总循环微粒、凝溶胶蛋白水平及其临床意义

目的 探讨子痫前期患者血浆可溶性内皮细胞蛋白C受体(s EPCR)、外周血总循环微粒(MPs)、凝溶胶蛋白(GSN)水平及其临床意义。方法 选取江苏省太仓市中医医院2021年3月至2023年3月建立产前登记并确诊的子痫前期孕妇103例作为病例组,同期建立产前登记且健康的孕妇200名作为对照组。根据病例组患者疾病严重程度将其分为轻度子痫前期组(71例)、重度子痫前期组(32例)。比较各组收缩压(SBP)、舒张压(DBP)、s EPCR、活化部分凝血酶时间(APTT)、凝血酶时间(TT)、凝血酶原时间(PT)、血小板计数(PLT)、外周血总循环MPs及GSN;sEPCR、外周血总循环MPs、GSN与患者血压水平的相关性采用Pearson相关分析。结果 病例组SBP、DBP高于对照组,PLT、TT、APTT低于对照组(P<0.05)。病例组的s EPCR、外周血总循环MPs高于对照组,GSN低于对照组(P<0.05)。重度子痫前期组SBP、DBP高于轻度子痫前期组,PLT低于轻度子痫前期组(P<0.05);两组TT、APTT比较,差异无统计学意义(P>0.05)。重度子痫前期组s EPCR、外周血总循环MPs高于轻度子痫前期组,GSN低于轻度子痫前期组(P<0.05)。子痫前期患者s EPCR、外周血总循环MPs与SBP、DBP呈正相关(r>0,P<0.05),GSN与SBP、DBP呈负相关(r<0,P<0.05)。结论 子痫前期患者s EPCR、外周血总循环MPs升高,GSN降低,并且与病情严重程度有关,可能与子痫前期患者呈高凝状态有一定的关系。

不同剂量替格瑞洛在急性心肌梗死PCI术后的应用效果分析

目的研究不同剂量替格瑞洛在急性心肌梗死经皮冠状动脉介入(PCI)术后的应用效果。方法选取2021年5月至2022年9月该院收治的100例行PCI术的急性心肌梗死患者为研究对象,根据治疗方案将患者分为常规组、研究组,每组50例。常规组PCI术后采用常规剂量替格瑞洛治疗,研究组PCI术后采用负荷剂量替格瑞洛治疗。比较术前、术后7 d两组TIMI血流分级、心功能指标[左室舒张末期内径(LVEDD)、左室射血分数(LVEF)、左室收缩末期内径(LVESD)]、血管内皮功能指标[循环内皮细胞(CEC)、血管性假血友病因子(vWF)、内皮素1(ET-1)、一氧化氮(NO)]及血清白细胞介素-8(IL-8)、白细胞介素-1β(IL-1β)、降钙素原(PCT)、粒巨噬细胞集落刺激因子受体α(CSF2RA)水平,比较两组不良心血管事件发生率。结果术后7 d研究组TIMI血流分级优于常规组(P<0.05)。术后7 d研究组LVESD、LVEDD小于常规组(P<0.05),LVEF高于常规组(P<0.05)。术后7 d研究组外周血CEC数量及血清vWF、ET-1水平低于常规组(P<0.05),血清NO水平高于常规组(P<0.05)。术后7 d研究组血清IL-8、IL-1β、PCT、CSF2RA水平低于常规组(P<0.05)。研究组不良心血管事件发生率[4.00%(2/50)]与常规组[8.00%(4/50)]相比,差异无统计学意义(P>0.05)。结论PCI术治疗急性心肌梗死患者后采取负荷剂量替格瑞洛治疗可有效调节患者TIMI血流分级,改善患者心功能及血管内皮功能,促进病情恢复,且未增加心血管不良事件发生风险。

急性白血病患者外周血中CECs、VEGF-C、VEGFR-2的变化及其临床意义

目的探讨循环内皮细胞(CECs)在急性白血病(AL)患者外周血中的数量变化及血清中血管内皮生长因子-C(VEGF-C)及其受体2(VEGFR-2)水平的变化及其临床意义.方法采用流式细胞仪(FCM)检测66例AL患者CECs的相对计数.采用ELISA的方法检测66例患者血清中VEGF-C、VEGFR-2的水平,并动态观察43例AL患者治疗前后的变化.结果 (1)AL患者初治组、CR组、NR组CECs外周血相对计数较对照组明显增高(P<0.05);CR组CECs相对计数较NR组明显减低(P<0.05);(2)AL患者初治组血清VEGF-C及VEGFR-2水平较对照组升高,2组差异有统计学意义(P<0.01);治疗后患者血清VEGF-C及VEGFR-2水平较治疗前降低,2组差异有统计学意义(P<0.01).结论 AL患者初治组外周血中CECs相对计数明显增高,治疗后CR组较NR组相对计数减降低;AL患者初治组血清中VEGF-C、VEGFR-2水平显著增高.CECs、VEGF-C、VEGFR-2其水平可能与AL患者病情发展、疗效及预后有一定关系.

Expression of platelet endothelial cell adhesion molecule-1 between pancreatic microcirculation and peripheral circulation in rats with acute edematous pancreatitis

OBJECTIVE: To study the ehanges of platelet endothelial cell adhesion molecule-1 (PECAM-1) expression on polymorphonuclear leukocytes (PMNs) in peripheral circulation anti pancreatic microcirculation in rats with acute edematous pancreatitis (AEP). METHODS: The model of AEP was established with 50 Wistar rats, and the changes of PECAM-1 expression on PMNs from the splenic vein and inferior vena cava were determined by flow cytometry. RESULTS: PECAM-I expression on PMNs showed no significant difference between pancreatic microcirculation and peripheral circulation at AEP2h and AEP4h time points. From the AEP4h to the AEP8h time point, PECAM-1 expression in peripheral circulation was up-regulated, but PECAM-1 expression in pancreatic microcirculation was down-regulated. PECAM-1 expression had a significant difference between pancreatic microcirculation and peripheral circulation at the AEP8h time point (P<0.05). CONCLUSION: PECAM-1 expression on PMNs is in a converse way between pancreatic microcirculation and peripheral circulation in AEP.

Tadalafil treatment had a modest effect on endothelial cell damage and repair ability markers in men with erectile dysfunction and vascular risk

The number of the circulating angiogenic cells （CACs） and colony forming units （CFUs） derived from cultured circulating mononuclear cells （MNCs） represents a laboratory surrogate for endothelial cell repair ability. The serum of men with erectile dysfunction （ED） and vascular risk factors （VRFs） showed an increased level of endothelial cell damage/dysfunction markers and reduced the numbers of CACs and CFUs derived from the cells of healthy men. We analyzed whether treating men with ED and VRFs with the selective phosphodiesterase type 5 inhibitor tadalafil improved the endothelial cell repair ability and reduced the levels of the serum markers of endothelial cell damage/dysfunction. MNCs from healthy men were cultured with 20% serum from 36 ED patients to obtain CACs and CFUs. The ED patients were evaluated before and after 4weeks of treatment with tadalafil （20 mg every other day） or with a placebo. The tadalafil treatment improved erectile function （P = 0.0028）, but had no effect on the inhibitory effects of serum from ED patients on the CACs and CFUs derived from healthy men. The levels of endothelin-1 （P = 0.011） and tissue type plasminogen activator （P = 0.005） were reduced after treatment compared to baseline and those of the placebo group, whereas no changes were observed in the E-selectin levels. The tadalafil treatment in the ED patients with VRFs resulted in only a modest effect on the laboratory measures of the endothelial cell damage/dysfunction and repair ability. The proposed beneficial effect of phosphodiesterase type 5 inhibition on vascular homeostasis requires further analysis.

Case Report: Pazopanib Treatment Response in a Patient with Metastatic Pleomorphic Dermal Sarcoma (Atypical Fibroxanthoma) with Circulating Tumor Cell-Derived Colonies as a Predictive Marker

Atypical fibroxanthomas (AFX) are rare skin tumors. These generally are superficial tumors, usually <3 cm red, fleshy, ulcerated skin lesions, that characteristically occur on sun-damaged skin, sometimes in immunocompromised or previously irradiated patients. These are part of a spectrum of more aggressive fibro-histiocytic neoplasms. In the older literature, these have been termed aggressive or metastatic AFX, but currently these have been reclassified as pleomorphic dermal sarcomas (PDS) and systemic undifferentiated pleomorphic sarcoma (UPS, formerly malignant fibrohistiocytic sarcoma, MFH). We present the case of a 64-year old woman who developed a deeply invasive PDS on the vertex of her scalp invading to the galea, with in-transit scalp metastases. Very little information is available about optimal treatment of metastatic PDS lesions. The patient was initially treated with 2 cycles of epirubicin/ifosfamide chemotherapy, resulting in life-threatening complications. A pretreatment peripheral blood sample was sent for CTC-derived colony assay. This sample grew 8 colonies from 10 ml blood. The tumor failed to respond to epirubicin and ifosfamide, and after several months of hospitalization, a second peripheral blood CTC-derived colony assay grew >376 colonies. The patient could not tolerate additional chemotherapy. She was therefore treated with the oral targeted agent pazopanib. The patient developed a dramatic biopsy-confirmed complete response. After 11 months of pazopanib treatment, a repeat CTC-derived culture sample grew only 8 colonies/10 ml blood. The complete response to pazopanib is still ongoing at over 41 months. To our knowledge, this is the first demonstration of clinical complete response of a PDS tumor following targeted therapy. An additional novel feature was the demonstration that CTC-derived colonies could be grown from the blood of a PDS patient. The number of colonies appeared to correlate with the clinical treatment response and seemed to function as a potential prognostic marker.

通督调神针刺结合化瘀通络灸治疗痰瘀阻络型后循环缺血性眩晕临床观察

目的观察通督调神针刺结合化瘀通络灸治疗痰瘀阻络型后循环缺血性眩晕(posterior circulation ischemic vertigo,PCIV)的临床疗效。方法将60例PCIV患者随机分为对照组和治疗组,每组30例。两组患者均接受基础治疗,治疗组患者再接受通督调神针刺和化瘀通络灸治疗,对照组患者再口服盐酸氟桂利嗪胶囊。治疗前后分别观察两组患者眩晕障碍量表(dizziness handicap inventory,DHI)评分、痰瘀阻络证积分、静脉血内皮祖细胞(endothelial progenitor cells,EPCs)计数,比较两组临床疗效。结果治疗组、对照组分别脱落3、2例患者。与治疗前比较,两组患者治疗后DHI评分、痰瘀阻络证积分均显著降低(P<0.05),EPCs计数显著升高(P<0.05),且治疗组DHI评分、痰瘀阻络证积分降低程度和EPCs计数升高程度均显著大于对照组(P<0.05)。治疗组临床疗效显著优于对照组(P<0.05)。结论通督调神针刺结合化瘀通络灸治疗PCIV的临床疗效确切。

AB027.Varying pattern of proteases secretion in Fuchs corneal endothelial dystrophy

Background:The goal of this project was to analyze the relationship between cell morphology and proteases/proteases inhibitors(PIs)secretion profile in fuchs endothelial corneal dystrophy(FECD)corneal endothelial cells(CECs).Methods:Cell morphology was determined using a circularity index(4π×area/perimeter2)for each CECs population extracted from surgical FECD specimens(N=2)and healthy Eye bank corneas(N=3).CECs were cultured 28 days post-confluency.Supernatant was collected and analysed using Proteome Profiler Array detecting 35 proteases and 32 PIs(R&D Systems).Proteome signal was analyzed using Image Studio Lite and correlated with the population’s circularity index.Results:Calculation of circularity index reported different morphologies among FECD populations(0.59±0.18 and 0.64±0.17)and healthy populations(0.44±0.18,0.66±0.13 and 0.71±0.11).Proteome arrays revealed the presence of 10 proteases(ADAMTS1,Cathepsin A,B,D,and X/Z/P,DPPIV/CD26,MMP-2,3 and 12,uPA/Urokinase)and 10 PIs(Protease Nexin II,Cystatin B and C,EMMPRIN/CD147,Latexin,Lipocalin-1,Serpin E1,TFPI,TFPI-2,TIMP-1,2 and 4).Healthy and FECD specimens showed similar variation patterns according to morphology for secretion of ADAMTS1,MMP-3 and 12.However,opposing patterns between healthy and FECD populations were observed for Cathepsin B and D.Moreover,some proteins did not show variation according to phenotype in healthy CECs,but did in FECD CECs:Cathepsin A,Cystatin C,TFPI-2 and total TIMPs.For the other proteins,secretion did not vary according to morphology or no specific pattern was distinguishable.Conclusions:To conclude,our results suggest that cell phenotype is linked to the secretion of certain proteases/PIs in both groups.However,there seems to be differences in secretion of particular proteases and PIs between FECD and healthy specimens as morphology did not have a similar influence.These differences might initiate an imbalance between proteases and PIs explaining the irregular thickening of the Descemet membrane seen in FECD.

微小RNA-124促进氧糖剥夺/复糖复氧后脑微血管内皮细胞血管新生的研究

目的探讨微小RNA(miR)-124对氧糖剥夺/复糖复氧(OGD/R)后脑微血管内皮细胞(BMEC)血管新生的作用及相关机制。方法将小鼠BMEC随机分为6组,正常对照组(con组),OGD/R组,OGD/R+过表达对照组(OGD/R+ov-con组),OGD/R+miR-124过表达组(OGD/R+ov-miR-124组),OGD/R+沉默对照组(OGD/R+si-con组),OGD/R+miR-124沉默组(OGD/R+si-miR-124组)。比较各组BMEC迁移距离、BMEC迁移数量、小管形成数量、CD31阳性细胞数、miR-124水平、15脂氧化酶(15-LOX)mRNA及蛋白表达水平。结果与con组比较,OGD/R组BMEC迁移距离、BMEC迁移数量、小管形成数量及miR-124水平降低(P<0.05,P<0.01);OGD/R组15-LOXmRNA及蛋白表达水平较con组显著升高(3.29±0.24vs1.00±0.00,4.31±0.39vs1.00±0.00,P<0.01)。与OGD/R+ov-con组比较,OGD/R+ov-miR-124组BMEC迁移距离、BMEC迁移数量、小管形成数量、CD31阳性细胞数、miR-124水平明显升高,15-LOXmRNA及蛋白表达水平降低(P<0.05);与OGD/R+si-con组比较,OGD/R+si-miR-124组BMEC迁移距离、BMEC迁移数量、小管形成数量、CD31阳性细胞数、miR-124水平降低,15-LOXmRNA及蛋白表达水平升高(P<0.05)。结论miR-124可能通过调控15-LOX水平促进OGD/R后BMEC血管新生,为缺血性脑卒中提供潜在治疗靶点。

糖尿病视网膜病变不同分期患者血清δ样蛋白4和环磷腺苷效应元件结合蛋白水平对微血管损伤的预测价值

目的分析糖尿病视网膜病变(DR)不同分期患者血清δ样蛋白4(DLL4)和环磷腺苷效应元件结合蛋白(CREB)水平对微血管损伤的预测价值。方法选择2020年2月—2023年2月收治的80例糖尿病患者为研究对象,依据DR不同分期分为无DR组(n=37)、非增生型DR组(n=19)和增生型DR组(n=24);检测血中DLL4及CREB水平,采用FACS Count流式细胞仪检测患者内皮祖细胞(EPC)、循环内皮细胞(CEC)及循环祖细胞(CPC)水平。结果增生型DR组血中DLL4及CREB水平最高,无DR组DLL4及CREB水平最低,差异有统计学意义(P<0.05);不同DR分期与DLL4、CREB水平呈显著正相关(P<0.05);糖尿病患者血中高DLL4及高CREB水平是影响DR分期的独立危险因素(P<0.05)。增生型DR组血中EPC、CPC水平最低,CEC水平最高,无DR组EPC、CPC水平最高,CEC水平最低,差异均有统计学意义(P<0.05);DLL4、CREB水平与EPC、CPC水平呈显著负相关(P<0.05),DLL4、CREB水平与CEC水平呈显著正相关(P<0.05);DLL4和CREB预测DR患者微血管损伤的曲线下面积(AUC)均大于0.85。结论随着DR分期增加,患者血清DLL4和CREB水平呈显著升高趋势,且DLL4和CREB对其微血管损伤具有较高的预测价值。

益气活血类中药促进血管新生的研究进展

血管新生是指在原有血管床的基础上再生出新的毛细血管,是防治缺血性疾病的重要方法。内皮细胞增殖、迁移与血管新生的发生发展密切相关。益气活血类中药单体及复方通过刺激内皮细胞增殖促进血管新生,改善局部缺血环境。因此文章综述了近年益气活血类中药单体、复方促进血管新生的研究成果,以期为中医药治疗缺血性疾病提供参考。

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.