Endothelial Specific Molecule-1 Research Articles

Functional analysis of ESM1 by shRNA-mediated knockdown of its expression in papillary thyroid cancer cells.

Endothelial specific molecule-1 (ESM1) is implicated as an oncogene in multiple human cancers. However, the function of ESM1 in papillary thyroid cancer (PTC) is not well understood. The current study aimed to investigate the effect of ESM1 on the growth, migration, and invasion of PTC to provide a novel perspective for PTC treatment. The expression levels of ESM1 in PTC tissues form 53 tumor tissue samples and 59 matching adjacent normal tissue samples were detected by immunohistochemical analysis. Knockdown of ESM1 expression in TPC-1 and SW579 cell lines was established to investigate its role in PTC. Moreover, cell proliferation, apoptosis, wound healing, and transwell assays were conducted in vitro to assess cell proliferation, migration and invasion. The findings revealed that ESM1 expression was significantly higher in PTC tissues than that found in paraneoplastic tissues (P<0.0001). Knockdown of ESM1 expression inhibited the proliferation, migration, and invasion of TPC-1 and SW579 cells in vitro. Compared with the control group, the mRNA and protein levels of ESM1 in PTC cells were significantly reduced following knockdown of its expression (P<0.01). In addition, ESM1-knockdown cells indicated decreased proliferation and decreased migratory and invasive activities (P<0.01, P<0.01, P<0.001, respectively). ESM1 was identified as a major gene in the occurrence and progression of PTC, which could increase the proliferation, migration, and invasion of PTC cells. It may be a promising diagnostic and therapeutic target gene.

ESM1 Interacts with c-Met to Promote Gastric Cancer Peritoneal Metastasis by Inducing Angiogenesis.

The peritoneum is the most common metastatic site of advanced gastric cancer and is associated with extremely poor prognosis. Endothelial-specific molecule 1 (ESM1) was found to be significantly associated with gastric cancer peritoneal metastasis (GCPM); however, the biological functions and molecular mechanisms of ESM1 in regulating GCPM remain unclear. Herein, we demonstrated that ESM1 expression was significantly upregulated in gastric cancer tissues and positively correlated with platelet endothelial cell adhesion molecule-1 (CD31) levels. Moreover, clinical validation, in in vitro and in vivo experiments, confirmed that ESM1 promoted gastric cancer angiogenesis, eventually promoting gastric cancer peritoneal metastasis. Mechanistically, ESM1 promoted tumor angiogenesis by binding to c-Met on the vascular endothelial cell membrane. In addition, our results confirmed that ESM1 upregulated VEGFA, HIF1α, and MMP9 expression and induced angiogenesis by activating the MAPK/ERK pathway. In conclusion, our findings identified the role of ESM1 in gastric cancer angiogenesis and GCPM, thus providing insights into the diagnosis and treatment of advanced gastric cancer.

P2Y2R‑mediated transactivation of VEGFR2 through Src phosphorylation is associated with ESM‑1 overexpression in radiotherapy‑resistant‑triple negative breast cancer cells.

We previously reported that radiotherapy‑resistant (RT‑R) triple negative breast cancer (TNBC) cells upregulate the expression of endothelial‑specific molecule‑1 (ESM‑1) compared with TNBC cells. In addition, ESM‑1 is involved in an increased proliferation and invasion of RT‑R‑TNBC cells compared with TNBC cells. It was further identified that, in RT‑R‑TNBC cells, P2Y2 purinergic receptor (P2Y2R)‑mediated activation of p21‑activated kinase1 (PAK1), protein kinase C (PKC), c‑Jun N‑terminal kinase (JNK) and p38 MAPKs is related to ESM‑1 expression via forkhead boxO1 (FoxO1) regulation. Notably, it has been reported that P2Y2R mediates the transactivation of vascular epithelial growth factor receptor2 (VEGFR2), and VEGFR2 is known to be involved in ESM‑1 expression. Therefore, in the present study, the involvement of VEGFR2 in the P2Y2R‑mediated ESM‑1 upregulation in RT‑R‑TNBC cells and the relationship between P2Y2R and VEGFR2 activation was further examined. Western blotting and reverse transcription‑PCR were used to monitor the expression of ESM‑1, and the results demonstrated that extracellular ATP treatment regulated the expression of ESM‑1 in a P2Y2R‑dependent manner in RT‑R‑MDA‑MB‑231 cells. In addition, extracellular ATP activated Src and VEGFR2 after 5 min of incubation, which was abolished by knockdown of P2Y2R expression. VEGFR2 activation in response to ATP was also decreased by inhibiting Src activity, suggesting that ATP‑activated P2Y2R regulates VEGFR2 phosphorylation via Src activation. Furthermore, ATP‑induced ESM‑1 expression was decreased by transfection with VEGFR2 small interfering RNA (siRNA). ESM‑1‑related signaling molecules, PAK1, PKC, JNK and p38 MAPKs, and the transcriptional regulator, FoxO1, which were activated by ATP, were also decreased following transfection with VEGFR2 siRNA. These results suggest that P2Y2R‑mediated transactivation of VEGFR2 through Src phosphorylation is associated with ESM‑1 overexpression in RT‑R‑TNBC cells.

Endocan as a potential marker in diagnosis and predicting disease severity in COVID-19 patients: a promising biomarker for patients with false-negative RT-PCR.

BackgroundEndothelial-specific molecule 1 (endocan) has emerged as an inflammatory biomarker in recent years. The purpose of this study was to investigate the diagnostic value of serum endocan levels in the prediction of COVID-19 disease among patients with a false-negative reverse transcription polymerase change reaction (RT-PCR) test, and also to determine its correlation with the clinical severity of the disease.MethodsThirty patients with positive RT-PCR results and 30 with false-negative RT-PCR results, both with suspected COVID-19 in terms of clinical, radiological, and laboratory findings, were included in the study. Thirty healthy controls were also enrolled.ResultsSerum endocan levels were estimated to be 821.8 ± 99.3 pg/mL in COVID-19 RT-PCR (+) patients, 803.9 ± 97.0 pg/mL in RT-PCR false (–) patients with suspected COVID-19, and 382.9 ± 37.5 pg/mL in the control group. No significant difference was observed between RT-PCR (+) and RT-PCR false (–) patients (P = 0.68). However, serum endocan levels differed significantly between patient groups and control group (P < 0.05). With a cut-off value of 444.2 pg/mL serum endocan levels differentiated COVID-19 cases from healthy individuals with 92% sensitivity and 80% specificity. Moreover, a significant positive correlation was observed between serum endocan levels and clinical severity (P < 0.01, r = 0.94).ConclusionsThere is a need for different laboratory markers capable of assisting diagnosis and showing COVID-19 infection in suspected COVID-19 RT-PCR false-negative patients. Endocan levels can be used as an assistant blood test for identifying COVID-19 patients with false-negative RT-PCR tests and in determining the clinical severity of the disease.

Circulating Endocan in Subjects with Obesity and Carbohydrate Disturbances

Objective: Endothelial-specific molecule 1 (endocan) is expressed by endothelial cells and may have a major role in the regulation of cell adhesion and in the pathogenesis of inflammatory disorders such as obesity and diabetes. Аim of this study is to evaluate endocan serum levels in patients with obesity and carbohydrate disturbances. Subjects and methods: The study group consisted of 163 patients with mean age 52,5±11,3 years, divided in four groups - obesity without carbohydrate disturbances, prediabetes, diabetes and 42 healthy individuals as a control group. Endocan levels were determined using a commercially available human enzyme-linked immune sorbent assay (ELISA) kit. Results: The levels of endocan were significantly higher in patients with prediabetes and obesity compared to the control group (618.92±874.29 ;425.85±391.76; vs.321.02±304.11pg/ml, p<0.05). Correlation analysis revealed that endocan correlates positively with triglyceride (TG) (r=0.219;p<0.05), gamma-glutamyl transferase (GGT) (r=0.225;p<0.05), visceral adiposity index (VAI) (r=0.186;p<0.05) and negative with HDL (r= -0.176;p<0.05). We found higher level of endocan in patients with three or more components of MS as well as higher levels of endocan were found in patients with dyslipidemia (p<0.05). ROC analyses determined circulating endocan levels to be of value for differentiating subjects with prediabetes (AUC=0.670,p=0.007, as a cut-off value of ≥183pg/ml had a sensitivity of 88% and specificity of 65%) Conclusion: The levels of endocan were increased in patients with obesity and carbohydrate disturbances. Further studies will elucidate the role of the biomarker in development of type 2 diabetes and its complications

Serum Endocan Levels on Admission Are Associated With Worse Clinical Outcomes in COVID-19 Patients: A Pilot Study.

Thrombotic and embolic complications in the cardiovascular system are evident and associated with worse prognosis in coronavirus disease 2019 (COVID-19) patients. Endothelial-specific molecule 1 (endocan) plays a role in vascular pathology. We hypothesized serum endocan levels on admission are associated with primary composite end point (mortality and intensive care unit hospitalization) in COVID-19 patients. Patients (n = 80) with laboratory, clinical, and radiological confirmed COVID-19 were included in this cross-sectional study. Ten milliliter of peripheral venous blood were drawn within 24 hours of admission to estimate serum endocan levels. Data were analyzed using SPSS version 26.0 (IBM). Patients with the primary composite end point had significantly higher serum endocan levels than patients without (852.2 ± 522.7 vs 550.2 ± 440.8 ng/L, respectively; P < .01). In the logistic regression analysis, only increased serum endocan levels and increase in age were independent predictors of the primary composite end point (P < .05). In the receiver operating characteristics curve analysis, we found that a serum endocan level of 276.4 ng/L had a 97% sensitivity and 85% specificity for prediction of the primary composite end point. Baseline serum endocan levels may prove useful as a prognostic factor in patients hospitalized for COVID-19.

Endothelial-Specific Molecule 1 (Endocan) as a Marker of Vascular Endothelial Regulation of Obesity-Associated Peripheral Polyneuropathy in the Non-Diabetic Obese Patients

Abstract Background: The increasing incidence of obesity and its co-morbid situations poses a great challenge to worldwide health. Obesity has numerous co-morbidities including airway illness, insulin resistance, type 2 diabetes, atherosclerosis, peripheral polyneuropathy (PN) and cancer. Endocan is a proteoglycan that could be used as a biomarker of endothelial dysfunction. Aim of Study: The current study aimed to investigate plasma endocan level in non-diabetic obese patients and to explore the association between circulatory endocan with the clinical and electrophysiological tests of PN in obese patients. Methods: This cross-sectional controlled study enrolled 170 obese patients and 100 control group. The obese group was sub-classified according to BMI (Body Mass Index) into three groups; all participants were subjected to a complete neurological examination. The motor nerve conduction study of [median nerve, ulnar nerve, and Common Peroneal Nerve (CPN)] and the sensory nerve conduction study of [median, ulnar and sural nerves] of all subjects were estimated. Blood sampling and biochemical analysis for parameters of metabolic syndrome (MetS) were done, in addition, Doppler evaluation of Carotid Intima Media Thickness (CIMT) using 0.9mm thickness as a cut-off point was used for identification of atherosclerosis. We measured plasma endocan by Enzyme-Linked Immunosorbent Assay (ELISA). Results: Obese patients with PN had statistically significant higher levels of plasma endocan (218.6±26.26) compared to obese patients without PN (135.5±21.6) and controls (13.1± 3.2). Plasma endocan level was positively correlated with Toronto Clinical Scoring System (TCSS) and negatively correlated with measures of electrophysiological tests; Motor Nerve Conduction Velocities (MNCV) of median, ulnar nerves, Sensory Nerve Conduction Velocities (SNCV) of median and ulnar nerves, Compound Muscle Action Potential (CMAP) amplitude of median and ulnar nerves and Sensory Nerve Action Potential (SNAP) amplitude of median, ulnar and sural nerves. The identification of optimum cut-off point of serum endocan could help in evaluating non-diabetic obese patients with PN in attempt to decrease health hazards related to neuropathy. Conclusion: Obese patients with PN had higher values of circulating endocan than obese patients without PN; the diagnostic power of circulating endocan was highly significant thus it could be used as a diagnostic biomarker of PN.

ESM1 as a Marker of Macrotrabecular-Massive Hepatocellular Carcinoma.

Macrotrabecular-massive hepatocellular carcinoma (MTM-HCC) is a novel morphological subtype of HCC associated with early relapse after resection or percutaneous ablation, independently of classical clinical and radiological prognostic factors. The aim of the present study was to identify immunohistochemical markers of MTM-HCC, to ease its diagnosis and implementation into clinical practice. To identify potential biomarkers of MTM-HCC, we first analyzed gene expression profiling data from The Cancer Genome Atlas study and further selected two candidate biomarkers. Performance of both biomarkers for diagnosis of MTM-HCC was further tested by immunohistochemistry in two independent series of 67 and 132 HCC biopsy samples. Analysis of RNA sequencing data showed that MTM-HCC was characterized by a high expression of neoangiogenesis-related genes. Two candidate biomarkers, Endothelial-Specific Molecule 1 (ESM1) and Carbonic Anhydrase IX (CAIX), were selected. In the discovery series, sensitivity and specificity of ESM1 expression by stromal endothelial cells for the detection of MTM-HCC were 97% (28/29), and 92% (35/38), respectively. Sensitivity and specificity of CAIX were 48% (14/29) and 89% (34/38). In the validation set, sensitivity and specificity of ESM1 for the identification of MTM-HCC were 93% (14/15) and 91% (107/117), respectively. Interobserver agreement for ESM1 assessment was good in both series (Cohen Kappa 0.77 and 0.76). Using a molecular-driven selection of biomarkers, we identified ESM1 as a reliable microenvironment immunohistochemical marker of MTM-HCC. The results represent a step toward the implementation of HCC morpho-molecular subtyping into clinical practice.

Endocan: a biomarker predicting successful reperfusion after coronary artery by-pass surgery of acute coronary syndrome patients.

Endothelial specific molecule 1 (Endocan) is a biomarker of the inflammatory process occurring in endothelial cells. It was shown that endocan was increased in acute coronary syndromes. We aimed at investigating the endocan levels in acute coronary syndrome patients after coronary bypass surgery. The study included 35 patients who presented with acute coronary syndrome to the Emergency Department. Serum endocan levels and other inflammatory markers were evaluated before and after coronary artery by-pass surgery. The correlation coefficients and their significance were calculated by Pearson's test and the difference in mean values before and after bypass surgery was calculated by paired sample t-test. Statistical analyses were performed using SPSS version 20.0 (SPSS Inc., Chicago, IL, USA). Statistical significance was taken as p<0.05. There was a significant change in serum hs-CRP and endocan levels and also LVEF before and after CABG operation (p<0.05). Serum endocan level was significantly decreased after successful reperfusion in our study. Thus, it may be an important biomarker in terms of clinical use as an indicator of successful reperfusion.

Endothelial nitric oxide synthase protein distribution and nitric oxide production in endothelial cells along the coronary vascular tree

ObjectiveFreshly isolated endothelial cells from both conduit arteries and microvasculature were used to test the hypothesis that eNOS protein content and nitric oxide production in coronary endothelial cells increases with vessel radius. MethodsPorcine hearts were obtained from a local abattoir. Large and small arteries as well as arterioles were dissected free of myocardium and homogenized as whole vessels. Additionally, endothelial cells were isolated from both conduit arteries and left ventricular myocardium by tissue digestion with collagenase, followed by endothelial cell isolation using biotinylated-anti-CD31 and streptavidin-coated paramagnetic beads. Purity of isolated endothelial cells was confirmed by immunofluorescence and immunoblot. ResultsIn whole vessel lysate, immunoblot analysis revealed that protein content for eNOS was greater in arterioles compared to small and large arteries. Nitric oxide metabolites (nitrite plus nitrate; NOx) levels measured from whole vessel lysate decreased as vessel size increased, with both arterioles and small arteries displaying significantly greater NOx content than conduit. Consistent with our hypothesis, both eNOS protein level and NOx were significantly greater in endothelial cells isolated from conduit arteries compared with those from coronary microvasculature. Furthermore, confocal microscopy revealed that eNOS protein was present in all conduit and microvascular endothelial cells, although eNOS staining was less intense in microvascular cells than those of conduit artery. ConclusionsThese findings demonstrate increased eNOS protein and NOx content in endothelial cells of conduit arteries compared with the microcirculation and underscore the importance of comparing endothelial-specific molecules in freshly isolated endothelial cells, rather than whole lysate of different sized vessels.

Clinical study of traditional Chinese medicine anti-inflammatory mixture for the microcirculation disturbance in patients with sepsis targetting at vascular endothelium

Objective To observe the clinical effect of traditional Chinese medicine inflammatory mixture on vascular endothelial cell injury, mcrocirculation disorder and organ dysfunction in sepsis. Methods 73 patients with sepsis were randomly divided into two groups: Anti-inflammatory mixture-treated group (40 cases) and control group (33 cases). Routine medicine therapy was given in both groups. Additionally, in the Anti-inflammatory mixture-treated group, 100 ml Anti-inflammatory mixture was given By oral or nasal feeding, every 12 hours for consecutive 7 days. Vascular endothelial injury index (Soluble thrombomodulin,Vascular endothelial growth factor-2, endothelial specific molecule-1), mcrocirculation disorder index (arterial blood lactate, central venous oxygen saturation, total vessel density, perfused vessel density, proportion of perfused vessels, microcirculatory flow index) of two group patients before and after therapy were observed and the sequential organ failure score (SOFA) was recorded before and after treatment. Results After treatment, the indexes of vascular endothelial injury sTM (12.37 ± 5.08 μg/L vs. 18.77 ± 6.88 μg/L, t=3.448), VEGF-2 [45.6 ng/L (14.3, 112.5) vs. 52.4 ng/L (17.2, 123.6), Z=4.009], ESM-1 (15.54 ± 4.09 ng/ml vs. 17.64 ± 6.79 ng/ml, t=-1.551), Lac (2.6 ± 1.2 mmol/L vs. 3.7 ± 1.8 mmol/L, t=4.115) and SOFA (4.1 ± 1.7 vs. 6.1 ± 3.2, t=-2.118) in anti-inflammatory mixture group decreased significantly than those in the control group. but the ScvO2 (0.719 ± 0.243 vs. 0.603 ± 0.201, t=-2.773), PVD (14.8 ± 5.8 mm/mm2vs. 13.1±5.1 mm/mm2, t=-5.114), PPV (59.1% ± 22.5% vs. 53.9% ± 20.6%, t=1.779), MFI (9.4 ± 4.6 vs. 7.2 ± 2.2, t=4.339) in the anti-inflammatory mixture group were significantly higher than those in the control group. Correlation analysis showed that the sTM and PPV were significantly negatively correlated (r=-0.875, P<0.01), ESM-1 was significantly negatively correlated with PVD, PPV and MFI (r=-0.877, P<0.01; r=-0.799, P<0.01; r=-0.821, P<0.01) and ESM-1 and SOFA were significantly positively correlated (r=0.840, P<0.01). Conclusions Anti-inflammatory mixture of Chinese medicine has a certain therapeutic effect on sepsis by protecting vascular endothelial cell function. Key words: Sepsis; Anti-inflammatory mixture; Compounds (TCD); Vascular endothelial cell; Microcirculatory disturbance; Clinical study

Inhibition of VEGF Signaling Reduces Diabetes-Exacerbated Brain Swelling, but Not Infarct Size, in Large Cerebral Infarction in Mice.

In light of repeated translational failures with preclinical neuroprotection-based strategies, this preclinical study reevaluates brain swelling as an important pathological event in diabetic stroke and investigates underlying mechanism of the comorbidity-enhanced brain edema formation. Type 2 (mild), type 1 (moderate), and mixed type 1/2 (severe) diabetic mice were subjected to transient focal ischemia. Infarct volume, brain swelling, and IgG extravasation were assessed at 3days post-stroke. Expression of vascular endothelial growth factor (VEGF)-A, endothelial-specific molecule-1 (Esm1), and the VEGF receptor 2 (VEGFR2) was determined in the ischemic brain. Additionally, SU5416, a VEGFR2 inhibitor, was treated in the type 1/2 diabetic mice, and stroke outcomes were determined. All diabetic groups displayed bigger infarct volume and brain swelling compared to nondiabetic mice, and the increased swelling was disproportionately larger relative to infarct enlargement. Diabetic conditions significantly increased VEGF-A, Esm1, and VEGFR2 expressions in the ischemic brain compared to nondiabetic mice. Notably, in diabetic mice, VEGFR2 mRNA levels were positively correlated with brain swelling, but not with infarct volume. Treatment with SU5416 in diabetic mice significantly reduced brain swelling. The study shows that brain swelling is a predominant pathological event in diabetic stroke and that an underlying event for diabetes-enhanced brain swelling includes the activation of VEGF signaling. This study suggests consideration of stroke therapies aiming at primarily reducing brain swelling for subjects with diabetes.

Tip-cell behavior is regulated by transcription factor FoxO1 under hypoxic conditions in developing mouse retinas.

Forkhead box protein O1 (FoxO1) is a transcription factor and a critical regulator of angiogenesis. Various environmental stimuli, including growth factors, nutrients, shear stress, oxidative stress and hypoxia, affect FoxO1 subcellular localization and strongly influence its transcriptional activity; however, FoxO1-localization patterns in endothelial cells (ECs) during development have not been clarified in vivo. Here, we reported that FoxO1 expression was observed in three layers of angiogenic vessels in developing mouse retinas and that among these layers, the front layer showed high levels of FoxO1 expression in the nuclei of most tip ECs. Because tip ECs migrate toward the avascular hypoxic area, we focused on hypoxia as a major stimulus regulating FoxO1 subcellular localization in tip cells. In cultured ECs, FoxO1 accumulated into the nucleus under hypoxic conditions, with hypoxia also inducing expression of tip-cell-specific genes, including endothelial-specific molecule 1 (ESM1), which was suppressed by FoxO1 knockdown. Additionally, in murine models, EC-specific FoxO1 deletion resulted in reduced ESM1 expression and suppressed tip-cell migration during angiogenesis. These findings indicated roles for FoxO1 in tip-cell migration and that its transcriptional activity is regulated by hypoxia.

Clinical study of therapeutic effect of Xuebijing injection on the treatment of vascular endothelial cell injury, microcirculation disorder and organ diysfunction in patients with sepsis

Objective To observe the clinical therapeutic effect of Xuebijing injection on vascular endothelial cell (VEC) injury, microcirculation disorder and organ dysfunction in patients with sepsis. Methods Seventy-three patients with sepsis were randomly divided into two groups: Xuebijing injection-treated group (40 cases) and control group (33 cases). Routine medicine therapy was applied in both groups.Additionally, the Xuebijing injection-treated group was treated with Xuebijing injection 100 ml and saline 100 ml by intravenous drip every 12 hours for consecutive 5 days.Vascular endothelial injury index, including soluble thrombomodulin(sTM), vascular endothelial growth factor 2 (VEGF-2), endothelial specific molecule 1 (ESM-1), microcirculation index of arterial blood lactic acid (Lac), central venous oxygen saturation (ScvO2), total microvessel density (TVD), the perfusion vascular density (PVD), proportion of perfused vessels (PPV) and microvessel flow index (MFI) of the two groups before and after therapy were observed and the sequential organ failure score (SOFA) was recorded before and after treatment. Results Senventy-three patients with sepsis had different degrees of increase in vascular endothelial damage markers, lactate and sequential organ failure scores in arterial blood, while the central venous blood oxygen saturation (ScvO2), the total vascular density of the sublingual microvasculature (TVD), perfused vessel density (PVD), proportion of perfused vessels (PPV) and microcirculatory flow index (MFI) decreased before treatment.After 5-day treatment, the above indicators of all patients were improved, the indexes in the Xuebijing injection group decreased significantly, compared with the control group , sTM ( (16.91±4.55) μg/L, (19.51±4.09) μg/L, t=-6.021, P<0.05), VEGF-2 (50.8(17.8, 127.7) ng/L vs.74.9(22.7, 155.1) ng/L, t=4.227, P<0.05), ESM-1 ( (10.20±2.43) μg/L vs.(14.80±3.52) μg/L, t=-4.113, P<0.05), Lac( (2.1±0.7) mmol/L vs.(3.7±1.1) mmol/L, t=2.366, P<0.05) and SOFA ( (5.9±2.1) vs.(8.7±2.6), t=-7.990, P<0.05). ScvO2 ( (0.771±0.153) % vs.(0.641±0.113) %, t=5.061, P<0.05), PVD ( (16.8±6.1) mm/mm2 vs.(12.1±5.1) mm/mm2, t=4.002, P<0.05), PPV ( (66.2±21.3) % vs.(50.4±19.3) %, t=-2.550, P<0.05) and MFI (6.2±2.4) vs.(3.8±2.2), t=-5.001, P<0.05) were significantly higher than those in the control group in the same period.sTM and PPV had a significant negative correlation (r=-0.755, P=0.000), PVD, PPV, ESM-1 and MFI were negatively correlated (r=-0.665, P=0.000; r=-0.600, P=0.000; r=-0.469, P=0.000), PPV, MFI and SOFA were negatively correlated (r=-0.798, P=0.000; r=-0.995, P=0.000); sTM, ESM-1 and SOFA were significantly positively correlated (r=0.883, P=0.000; r=0.881, P=0.000). Conclusion Vascular endothelial cell dysfunction probably plays an important role in the pathophysiology of sepsis and Xuebijing injection has therapeutic effect on sepsis by protecting vascular endothelial cell function. Key words: Sepsis; Xuebijing injection; Vascular endothelial cell; Microcirculation disorder

Significance of serum endothelial cell specific molecule-1 (Endocan) level in patients with erectile dysfunction: a pilot study.

This study aimed to measure the serum endocan level of patients with erectile dysfunction (ED) and to investigate the possible association between the Endothelial-specific molecule-1 (Endocan) level and ED. Twenty healthy and sixty-four male patients included in the study were divided into four groups: severe ED (19 patients), moderate ED (24 patients), mild ED (21 patients) and control group (20 healthy men). The erectile function of all the patients was evaluated using the International Index of Erectile Function-5 (IIEF-5) questionnaire. The body mass index (BMI) of each participant was determined, together with levels of fasting blood glucose, total testosterone, low- and high-density lipoprotein cholesterol, triglyceride and endocan in serum samples. No significant difference was found between the three ED groups and the control group in terms of the mean age, BMI and the levels of cholesterol and fasting blood glucose (P>0.05). The mean serum endocan level was found to 1.076±0.5, 0.674±0.40 and 0.671±0.3 ng ml-1 in the severe, moderate and mild ED groups, respectively. This indicated that the highest value was obtained from the severe ED group, and the difference between the severe ED group and the other groups was statistically significant. In the control group, the serum endocan level was 0.73±0.46 ng ml-1, which was significantly higher compared to the moderate and mild ED groups (P<0.05). The significant difference between the control and ED groups in terms of the serum endocan level can assist in the evaluation of endothelial pathologies in the etiology ED.

Endothelial Specific Molecule 1 (Esm‐1) Inhibits Macrophage Infiltration in a Mouse Model of Goodpasture's Syndrome

Goodpasture's syndrome is an autoimmune disorder caused by pathogenic antibodies to kidney and lung basement membranes causing acute kidney and lung injury, respectively. We modeled this syndrome by treatment with anti‐GBM (Glomerular Basement Membrane) antibodies leading to glomerular and tubulointerstitial inflammation in mice. Endothelial Specific Molecule1 (Esm‐1)/endocan is a secreted glycoprotein highly expressed in kidney glomeruli and lung endothelium and is proposed to disrupt molecular interactions that mediate leukocyte infiltration and anti‐GBM nephritis. To discover the in vivo functional role of Esm‐1 in leukocyte infiltration in anti‐GBM mice, we delivered Esm‐1 plasmid for stable integration to anti‐GBM mice and controls by hydrodynamic tail vein injection. We quantified kidney leukocyte infiltration by separating glomerular and tubulointerstitial compartments, dissociating into single cells, and measuring leukocyte subtypes by flow cytometry. In contrast to i.p. administration of recombinant Esm‐1 which has an abbreviated half‐life in mouse serum (less than 2 hours), we induced ~12‐fold increase in circulating Esm‐1 that persists for 2 weeks compared to control injected mice, by hydrodynamic injection of an Esm‐1‐expressing plasmid (11242 +/− 3324 vs. 958 +/− 188 pg/mL, p&lt;0.05, N=3–5/group). We demonstrate that by flow cytometry, anti‐GBM mice show significantly increased glomerular (3.4 +/− 0.47% vs. 1.1 +/− 0.33%, p &lt; 0.01, N=8/group) and tubulointerstitial (1.4 +/− 0.14% vs. 0.1+/− 0.032%, p &lt; 0.01, N=8/group) CD45+ leukocytes seven days post treatment with anti‐GBM antisera. The increase in CD45+ cells includes increased glomerular (3.4 +/− 0.47% vs. 1.1 +/− 0.33%, p &lt; 0.01) and tubular (1.2 +/− 0.14% vs. 0.1+/− 0.025%, p &lt; 0.01) macrophages. By increasing circulating Esm‐1, we significantly decreased macrophage infiltration in the tubulointerstitium (4.2 ± 1.3% vs. 1.6 ± 0.4%, P &lt; 0.01), but did not influence glomerular macrophage infiltration (2.3 ± 1.2% vs. 1.5 ± 0.2%, P = 0.13). Furthermore, Esm‐1 does not alter kidney neutrophils (glomerular: 0.013 ± 0.007% vs. 0.010 ± 0.006%, p = 0.61; tubulointerstitial: 0.19 ± 0.02% vs. 0.27 ± 0.07%, p = 0.085) or lymphocytes(glomerular: 1.7 ± 0.8% vs. 1.9 ± 0.9%, p = 0.89; tubulointerstitial: 1.9 ± 0.70% vs. 2.2 ± 0.71%, p = 0.64). We also found that inhibition of macrophage infiltration was dependent on glycosylation of Esm‐1. Furthermore, we have generated Esm‐1 knockout mice for complementary studies. We conclude that in contrast to deletion of the inflammatory mediator ICAM‐1, Esm‐1 inhibits inflammation in a tissue and cell‐specific manner. Esm‐1 provides a natural biologic inhibitor of inflammation, and based on its expression pattern, may be a key therapeutic target in pulmonary‐renal syndromes.Support or Funding Information1. Larry L. Hillblom Foundation2. Holmgren Family3. NIDDK

Endocan Levels and Subclinical Atherosclerosis in Patients With Systemic Lupus Erythematosus.

Systemic lupus erythematosus (SLE) is a chronic autoimmune disease of unknown etiology. A major cause of morbidity and mortality in SLE is accelerated atherosclerosis. Endothelial-specific molecule 1 (endocan) is a potential predictor of vascular events and is expressed in response to inflammatory cytokines in endothelial cells. We investigated the relationship between endocan and carotid intima-media thickness (cIMT) as a marker of early atherosclerosis. We included 44 women with SLE and 44 healthy women as controls. Disease severity of SLE was evaluated using the SLE Disease Activity Index. Endocan, C-reactive protein, erythrocyte sedimentation rate (ESR), and lipid panel were measured. The cIMT was 0.70 (range: 0.45-1.20) mm in patients with SLE and 0.40 (0.25-0.60) mm in controls (P < .001). Endocan value was 1.6 ± 0.9 ng/mL in controls and 2.2 ± 1.0 ng/mL in patients with SLE (P = .014). Endocan levels were positively correlated with cIMT (r = .469, P < .001), body mass index (r = .373, P = .013), and ESR (r = .393, P = .008). Endocan level may be associated with subclinical atherosclerosis in SLE. Consequently, endocan levels may be a promising clinical tool for patients with SLE as a guide for preventive strategy.

Abstract TMP57: Vascular Endothelial Growth Factor Signaling Contributes to Comorbidity-enhanced Edema in Ischemic Stroke

Introduction: Stroke-induced brain edema is a life-threatening complication for patient. Pre-clinical studies have shown that comorbidities such as diabetes (DB) and hyperlipidemia (HL) are associated with the enhanced edema, however the underlying mechanism is not clear. Vascular endothelial growth factor (VEGF)-A has a major role in angiogenesis and vascular permeability by binding with the VEGF receptor 2 (VEGFR-2). Hypothesis: We therefore hypothesized that VEGF signaling contributes to comorbidity-enhanced edema in ischemic stroke. Methods: DB was induced in male C57BL/6 mice by feeding a high fat diet for 8 weeks with or without streptozotocin administration. HL was generated in male ApoE knock-out mice by feeding a high fat diet for 8 weeks. The mice were then subjected to 30 min transient middle cerebral artery occlusion and brain infarct and edema size were determined at 3 days post-stroke. Gene or protein expressions of VEGF-A, VEGFR-2, and the downstream pathway (phospho(p)-Akt and endothelial specific molecule (Esm)-1) were determined in the brain following stroke. Results: DB and HL mice displayed hyperglycemia and hyperlipidemia. Following stroke, DB and HL mice exhibited larger infarct and edema, and increased VEGF-A and VEGFR-2 expression compared to normal mice. The increase was accompanied by increased pAkt and Esm1 expression. We also found that the VEGFR-2 gene was positively correlated with the severity of edema in both comorbid conditions but not in normal condition (NC) (Fig. A and B). Conclusion: The results demonstrate that the VEGF signaling may underlie for comorbidity-enhanced brain edema in stroke. A treatment strategy aimed at reducing edema by targeting VEGF signaling is suggested for stroke patients with the comorbidities.

Serum Endocan Level and the Severity of Coronary Artery Disease: A Pilot Study.

Endothelial-specific molecule 1 (endocan) is expressed in endothelial cells. We investigated the relationship between acute coronary syndrome (ACS) and serum endocan levels. We included 30 individuals as a control group and 53 patients diagnosed with ACS. The severity of coronary artery disease was assessed by a modified Gensini stenosis and SYNTAX scoring system. There was a significant difference in serum endocan levels between the control group and the ACS group (0.75 ± 0.13 vs 0.86 ± 0.25 ng/mL, P = .014). There was also a significant difference in serum endocan levels between diabetic patients with ACS and nondiabetic patients with ACS (1.02 ± 0.33 vs 0.81 ± 0.21 ng/mL, P = .016). There was no significant correlation between serum endocan level, Gensini, and SYNTAX score (r = .11, P = .53 and r = .16, P = .37). Endocan, a new biomarker of endothelial pathology, is significantly increased in patients with ACS.

Model microgravity enhances endothelium differentiation of mesenchymal stem cells

Mesenchymal stem cells (MSCs) are capable of differentiation into multilineage cell types under certain induction conditions. Previous studies have demonstrated that physical environments and mechanical force can influence MSC fate, indicating that these factors may be favorable inducers for clinical treatment. Our previous study found that MSCs are spread with a spindle shape when cultured in normal gravity (NG), and under modeled microgravity (MMG) for 72h, they become unspread and round and their cytoskeleton fibers are reorganized. These morphological changes affected the function of MSCs through the activity of RhoA. We examined the responses of MSCs under MMG stimulation, followed with VEGF differentiation. We found that MSCs under MMG for 72h were differentiated into endothelial-like cells by detecting the expression of endothelial-specific molecules (Flk-1 and vWF), which were also able to form a capillary network. Their endothelial differentiation potential was improved under MMG compared with that under NG. We believe that this method is a novel choice of MMG stimulation for neovascularization. This phenomenon may increase the potential of MSC differentiation, which might be a new strategy for the treatment of various vascular diseases and improve vascularization in tissue engineering.