Association of Myostatin With Complications and Cognition in Lung Cancer Patients With Sarcopenia.

INTRODUCTION: The risk of surgery and postoperative complications increases greatly in frail older patients with sarcopenia. The purpose of this study is to explore the correlation between myostatin (MSTN) levels and cognitive function and postoperative pulmonary complications (PPCs) in older patients undergoing thoracoscopic lobectomy and to determine whether MSTN could be used to predict the risk of postoperative complications and cognitive impairment. METHODS: A prospective observational study was conducted at the First Affiliated Hospital of Bengbu Medical College, China, between January 2023 and June 2023. The risk factors of PPCs and postoperative cognitive impairment were studied using backward stepwise logistic regression analysis. The independent factors were formed into a linear regression equation to construct a risk score model for each patient. The 122 patients who participated in the study were divided into two groups, a low-level group and a high-level group, based on an MSTN level cut-off; the preoperative MSTN cut-off values was 25.55 ng/mL for cognitive dysfunction and 22.29 ng/mL for PPCs. The PPCs and cognitive function of the groups were compared. RESULTS: Preoperative MSTN was confirmed as a risk factor for postoperative cognitive dysfunction and PPCs. After surgery, the proportion of patients with cognitive impairment in the high-level group was significantly higher than in the low-level group (P < 0.001). In the high-level group, the incidence of respiratory tract infections was 17.9% higher (P = 0.021), hypoxaemia was 20.5% higher (P = 0.001) and respiratory failure was 14.4% higher (P = 0.012) than in the low-level group. In addition, a high level of MSTN increased the length of hospital stay (P < 0.001) and decreased the Barthel Index score (P < 0.001). CONCLUSIONS: The study findings suggest that MSTN could be used as an index to predict complications and cognitive impairment after thoracoscopic lobectomy in older patients with sarcopenia and to provide evidence for reducing postoperative cognitive impairment and PPCs.

Predicting the Risk of Postoperative Delirium in Elderly Patients Undergoing Hip Arthroplasty: Development and Assessment of a Novel Nomogram.

OBJECTIVE: To construct and internally validate a nomogram that predicts the likelihood of postoperative delirium in a cohort of elderly individuals undergoing hip arthroplasty. METHODS: Data for a total of 681 elderly patients underwent hip arthroplasty were retrospectively collected and divided into a model (n = 477) and a validation cohort (n = 204) according to the principle of 7:3 distribution temporally. The assessment of postoperative cognitive function was conducted through the utilization of The Confusion Assessment Method (CAM). The nomogram model for postoperative cognitive impairments was established by a combination of Lasso regression and logistic regression. The receiver operating characteristic (ROC) curve, calibration plot, and decision curve analysis (DCA) were used to evaluate the performance. RESULTS: The nomogram utilized various predictors, including age, body mass index (BMI), education, preoperative Barthel Index, preoperative hemoglobin level, history of diabetes, and history of cerebrovascular disease, to forecast the likelihood of postoperative delirium in patients. The area under the ROC curves (AUC) for the nomogram, incorporating the aforementioned predictors, was 0.836 (95% CI: 0.797-0.875) for the training set and 0.817 (95% CI: 0.755-0.880) for the validation set. The calibration curves for both sets indicated a good agreement between the nomogram's predictions and the actual probabilities. CONCLUSION: The use of this novel nomogram can help clinicians predict the likelihood of delirium after hip arthroplasty in elderly patients and help prevent and manage it in advance.

Dexmedetomidine mitigates acute kidney injury after coronary artery bypass grafting: a prospective clinical trial.

INTRODUCTION AND OBJECTIVES: To evaluate the impact of dexmedetomidine impact on cardiac surgery-associated acute kidney injury (CSA-AKI), kidney function, and metabolic and oxidative stress in patients undergoing coronary artery bypass grafting with heart-lung machine support. METHODS: A randomized double-masked trial with 238 participants (50-75 years) undergoing coronary artery bypass grafting was conducted from January 2021 to December 2022. The participants were divided into Dex (n=119) and NS (n = 119) groups. Dex was administered at 0.5 mcg/kg over 10minutes, then 0.4 mcg/kg/h until the end of surgery; the NS group received equivalent saline. Blood and urine were sampled at various time points pre- and postsurgery. The primary outcome measure was the incidence of CSA-AKI, defined as the occurrence of AKI within 96hours after surgery. RESULTS: The incidence of CSA-AKI was significantly lower in the Dex group than in the NS group (18.26% vs 32.46%; P=.014). Substantial increases were found in estimated glomerular filtration rate value at T4-T6 (P<.05) and urine volume 24hours after surgery (P<.01). Marked decreases were found in serum creatinine level, blood glucose level at T1-T2 (P<.01), blood urea nitrogen level at T3-T6 (P<.01), free fatty acid level at T2-T3 (P<.01), and lactate level at T3-T4 (P<.01). CONCLUSIONS: Dex reduces CSA-AKI, potentially by regulating metabolic disorders and reducing oxidative stress. Registered with the Chinese Clinical Study Registry (No. ChiCTR2100051804).

Cancer-associated fibroblasts: from basic science to anticancer therapy.

Cancer-associated fibroblasts (CAFs), as a central component of the tumor microenvironment in primary and metastatic tumors, profoundly influence the behavior of cancer cells and are involved in cancer progression through extensive interactions with cancer cells and other stromal cells. Furthermore, the innate versatility and plasticity of CAFs allow their education by cancer cells, resulting in dynamic alterations in stromal fibroblast populations in a context-dependent manner, which highlights the importance of precise assessment of CAF phenotypical and functional heterogeneity. In this review, we summarize the proposed origins and heterogeneity of CAFs as well as the molecular mechanisms regulating the diversity of CAF subpopulations. We also discuss current strategies to selectively target tumor-promoting CAFs, providing insights and perspectives for future research and clinical studies involving stromal targeting.

Depressive symptoms increase the risk of falls and injurious falls in Chinese adults aged ≥ 45 years: A 3-year cohort study.

Background: Falls and depressive symptoms are both public health concerns in China, but the effects of depressive symptoms on falls and injurious falls have not been thoroughly investigated. Methods: This population-based prospective cohort study used data derived from adults aged ≥45 years acquired from the 2015 and 2018 China Health and Retirement Longitudinal Study. Data were analyzed from August 2021 to December 2021. Self-reported depressive symptoms were determined using a 10-item Center for Epidemiologic Studies Depression scale (CESD-10) with a total score range of 0-30. Item responses of 3-4 or 5-7 days were deemed indicative of specific depressive symptoms. The outcome variables were self-reported accidental falls and injurious falls. Results: Of the 12,392 participants included in the study, 3,671 (29.6%) had high baseline depressive symptoms (CESD-10 scores ≥ 10), 1,892 (15.3%) experienced falls, and 805 (6.5%) experienced injurious falls during 2015-2018 follow-up. High depressive symptoms increased the risk of falls [odds ratio (OR) 1.34, 95% confidence interval (CI) 1.19-1.50] and injurious falls (OR 1.28, 95% CI 1.09-1.51) in a multivariable logistic regression model adjusted for major demographic, health-related, and anthropometric covariates. All of the 10 specific depressive symptoms except "felt hopeless" were associated with falls, and four specific symptoms significantly increased the risk of injurious falls; "had trouble concentrating" (OR 1.32, 95% CI 1.13-1.55); "felt depressed" (OR 1.32, 95% CI 1.12-1.55); "everything was an effort" (OR 1.23, 95% CI 1.04-1.45); and "restless sleep" (OR 1.18, 95% CI 1.02-1.40). Conclusion: High depressive symptoms are significantly related to risk of falls and injurious falls. Four specific symptoms (had trouble concentrating, felt depressed, everything was an effort, and restless sleep) increase the risk of injurious falls in Chinese adults aged ≥ 45 years.

Bioassay-guided isolation of anti-tumor polyprenylphloroglucinols from Calophyllum polyanthum and primary mechanism.

Five compounds were isolated from Calophyllum polyanthum leaves (10.09 g) by bioassay-guided fractionation to evaluate their anti-tumor activity. Among these compounds, apetalic acid (1) demonstrated significant inhibitory activity against 8 types of tumor cells (MHCC97H, CNE1, CNE2, B16, LOVO, SW480, A549, 1299), especially against two colon cancer cells (LOVO, SW480). Apetalic acid could inhibit cell proliferation, migration, invasion and induce apoptosis. It could significantly up-regulate the expression levels of apoptosis-related genes (BAX, Caspase-9,) and proteins (BAX, Cleaved-caspase-9, Cleaved-caspase-3) and down-regulated the expression of inhibitor of apoptosis gene (Bcl-2) and proteins (Bcl-2, phosphorylated AKT). Possible mechanism of the antitumor activity of apetalic acid derived from Calophyllum polyanthum supports its use in the prevention and treatment of colorectal cancer.

Ononin alleviates H2O2-induced cardiomyocyte apoptosis and improves cardiac function by activating the AMPK/mTOR/autophagy pathway.

Ononin (ON) is an isoflavone with numerous reported bioactivities, including anti-oxidative, anti-inflammatory and neuroprotective effects. Autophagy is a critical homeostatic process in the body that has been reported to closely associate with the apoptotic processes of cardiomyocytes. Using flow cytometry, western blotting, echocardiography and Masson's staining, the present study investigated the effects of ON on H2O2-induced cardiomyocyte apoptosis and myocardial infarction, in addition to any potential underlying molecular mechanisms. H2O2 treatment reliably induced apoptosis in H9C2 cells. The anti-apoptotic effects of ON were revealed by flow cytometry results and by the downregulation of cleaved-caspase 3. Further investigations indicated that ON may alleviate apoptosis by enhancing autophagy, as evidenced by increased microtubule-associated proteins 1A/1B light chain 3B expression and p62 degradation. Activation of the 5' AMP-activated protein kinase (AMPK)/mTOR pathway was observed after ON administration following H2O2-induced cardiomyocyte injury. However, these anti-apoptotic effects mediated by ON were lost after autophagy inhibition by chloroquine or AMPK inhibition by Compound C. Finally, the protective effects of ON on cardiomyocytes in vitro could also be observed in vivo. A myocardial infarction model was established by ligating the left anterior descending branch of the rat heart. Using echocardiography and Masson's staining, ON was shown to increase the ejection fraction and decrease cardiac fibrosis in rats with myocardial infarction. These results suggest that ON exerts cardioprotective effects by improving autophagy via the AMPK/mTOR signaling pathway.

Selective catalytic degradation of a lignin model compound into phenol over transition metal sulfates.

Transition metal salts were employed as the catalysts to improve the selective degradation of the α-O-4 lignin model compound (benzyl phenyl ether (BPE)) in the solvothermal system. The results concluded that most of the transition metal salts could enhance BPE degradation. Among which, NiSO4·6H2O exhibited the highest performance on BPE degradation (90.8%) for 5 h and phenol selectivity (53%) for 4 h at 200 °C. In addition, the GC-MS analysis indicated that the intermediates during BPE degradation included a series of aromatic compounds, such as phenol, benzyl methyl ether and benzyl alcohol. Furthermore, the mechanisms for BPE degradation and phenol selectivity in the NiSO4·6H2O system involved the synergetic effects between the acid catalysis and coordination catalysis, which caused the effective and selective cleavage of the C-O bonds.

[Differential screening and functional prediction analysis of miRNA expression profiles in periodontitis].

PURPOSE: To detect the gene expression of miRNAs in patients with periodontitis and to explore their biological functions and involved signaling pathways. METHODS: Bioinformatics analysis of gene chip data from 158 periodontitis patients and 40 healthy controls of the microarray database GSE54710 were performed. The expression changes of miRNAs were analyzed. The involved biological function and signal path was predicted. SPSS 19.0 software package was used for statistical analysis. RESULTS: Five miRNAs (hsa-miR-451, hsa-miR-223, hsa-miR-486-5p, hsa-miR-3917, hsa-miR-671-5p) were significantly up-regulated, and 4 miRNAs (hsa-miR-203, hsa-miR-210, hsa-miR-1246, hsa-miR-1260) were significantly down-regulated. Among them, there were 584 target genes of hsa-miR-1260 and 139 target genes of hsa-miR-451. KEGG pathway enrichment analysis showed that hsa-miR-1260 target gene was significantly enriched into 12 signaling pathways such as TGF-beta, and hsa-miR-451 target gene was significantly enriched into 17 signaling pathways. CONCLUSIONS: miRNAs expression profiles were obtained in periodontitis tissues, periodontitis-induced hsa-miR-1260 and hsa-miR-451 may play a key role in the pathophysiology of periodontitis.

Long noncoding RNA homeobox A11 antisense promotes transforming growth factor ß1­induced fibrogenesis in cardiac fibroblasts.

Cardiac fibrosis is closely associated with various heart diseases and is an important pathological feature of cardiac remodeling. However, detailed mechanisms underlying cardiac fibrosis remain largely unknown. Long noncoding RNAs (lncRNAs) are reported to serve significant roles in the development of cardiac fibrosis. The present study aimed to identify the role of a novel lncRNA, homeobox A11 antisense (HOXA11­AS), in cardiac fibrosis. Overexpression of HOXA11­AS in mouse cardiac fibroblasts (CFs) increased the expression of transforming growth factor ß1 (TGFß1) and its downstream molecules, while knockdown of HOXA11­AS inhibited the TGFß1 signaling pathway. Furthermore, as determined by colony formation and MTT assays, HOXA11­AS overexpression promoted colony formation and viability in mouse CFs, while HOXA11­AS knockdown had the opposite effect. In addition, overexpression of HOXA11­AS increased cell migration and invasion in the Transwell assays, whereas expression knockdown decreased the metastatic ability of cells. In order to explore the detailed mechanism, co­transfection of HOXA11­AS expression plasmid and siTGFß1 into CFs resulted in increased cell proliferative rate and cell metastasis through the TGFß1 signaling pathway. Taken together, the present study suggested that the lncRNA HOXA11­AS may be a potential therapeutic target against cardiac fibrosis, and provided a novel insight into the diagnosis and treatment of clinical cardiac fibrosis.

A rare case of B-lymphoproliferative disorder with villous lymphocytes harboring t(8;14)(q24;q32) translocation.

Splenic lymphoma with villous lymphocytes (SLVL) or splenic marginal zone lymphoma with circulating villous lymphocytes is rare, and prolymphocytic transformation of SLVL is rarer. At present, only one case of SLVL with t(8;14)(q24;q32) translocation has been reported. In this study, we report a case of B-lymphoproliferative disorder with villous lymphocytes harboring t(8;14)(q24;q32) chromosome translocation that we inclined to SLVL with a prolymphocytic transformation. A 73-year-old female showed marked hepatosplenomegaly and high lymphocytosis (lymphocytes > 200 × 109/L). The abnormal lymphocytes had short coarse villi and round nuclei with prominent nucleoli. The immunophenotypes showed CD19+, CD20+, HLA-DR+, CD22+, CD5+, Kappa+, CD25dim, CD71dim, Lambda-, CD7-, CD10-, CD23-, CD34-, CD33-, CD13-, CD14-, CD117-, CD64-, CD103-, and CD11c-. The karyotype showed complex abnormality: 46XX,+ 3,-10, t(8;14)(q24; q32)[11]/46XX[9]. The cytoplasmic projection, immunological characteristics, and trisomy 3 chromosome abnormality supported the diagnosis of SLVL. However, the presence of prominent nucleoli and high lymphocytosis suggested prolymphocytic transformation, probably as a result of t(8,14) chromosome translocation. In this report, we described an unusual case of B-lymphoproliferative disorder with villous lymphocytes harboring t(8;14)(q24;q32) translocation, which could provide help in the diagnosis and differential diagnosis of B-lymphocytic proliferative diseases.

Cryptotanshinone inhibits VEGF-induced angiogenesis by targeting the VEGFR2 signaling pathway.

OBJECTIVE: Anti-angiogenesis has been proposed as an important strategy for angiogenesis-related diseases. Cryptotanshinone (CPT), an active component of Salvia miltiorrhiza, may be a potential inhibitor of angiogenesis. However, the molecular mechanisms underlying its anti-angiogenic activities remain poorly understood. This study is to investigate the effects of CPT on VEGF-induced angiogenesis and VEGFR2 signaling pathway in human umbilical vein endothelial cells (HUVECs). METHODS: HUVECs were treated with different concentration of CPT (5-20µmol/L) and the viability, endothelial cell migration, invasion, and tubular-like structure formation of HUVECs were detected by MTT, wound-healing migration, Transwell invasion and Matrigel tube formation assays, respectively. To assess the effect of CPT on VEGFR2 signaling pathway, VEGF-induced phosphorylation of VEGFR2 and its downstream molecules, including ERK1/2, p90RSK, Src and FAK were analyzed by Western blot. RESULTS: CPT significantly suppressed VEGF-induced cells proliferation, migration, invasion, and tubular-like structure formation in HUVECs in a dose- and time-dependent manner. Western blot results revealed that CPT significantly suppressed VEGF-induced phosphorylation of VEGFR2 and its key downstream protein kinases, including p-ERK1/2, p-p90RSK, pY416-Src and pY576/577-FAK, which are responsible for endothelial cell migration, proliferation, and survival. CONCLUSION: Our study suggested that CPT potently inhibits VEGF-induced angiogenesis by suppressing VEGFR2 activation and its downstream Src/FAK and ERK1/2 signaling pathways in HUVECs, highlighting the therapeutic potential for the treatment of angiogenesis-related diseases.

Embedded structure fiber-optic radiation dosimeter for radiotherapy applications.

An investigation into a novel in-vivo PMMA (polymethyl methacrylate) plastic fiber-optic dosimeter for monitoring low doses of ionizing radiotherapy radiation in real time and for integrating measurements is presented. The fabricated optical fiber tip possessed an embedded structure. A scintillation material, terbium-doped gadolinium oxysulfide (Gd2O2S:Tb), capable of emitting visible light at around 545 nm which is ideal for transmission through the PMMA when exposed to ionizing radiation was embedded in the PMMA plastic fiber. The dose rate of incident ionizing radiation is measured by analyzing the signal intensity emitted from the scintillation material which propagates through the fiber to a distal MPPC (multi-pixel photon counter). The dosimeter exhibits good repeatability with an excellent linear relationship between the fiber-optic dosimeter output and the absorbed radiation dose with an outstanding isotropic response in its radial angular dependence.

[Meta-analysis on effect of Shenfu injection in treating angina pectoris].

To systematically evaluate the efficiency and safety of Shenfu injection in treating patients with angina pectoris. Retrievals were made in Embase, Pubmed, Cochrane Library, Clinical Trials.gov, CNKI, CBM, VIP and Wanfang (before September 2015) for randomized or semi-randomized controlled trials reporting data of Shenfu injection in the adjuvant treatment of angina pectoris. The quality of included trials was evaluated according to tool evaluation at cochrane.org. STATA version 12.0 was applied for Meta analysis after quality assessment of included studies. Finally, a total of 17 studies, including 16 randomized controlled trials (RCTs) and 1 controlled clinical trial (CCT) involving 1 309 patients, met the inclusion criteria, of which 659 patients received Shenfu injection treatment. Meta-analysis results showed that Shenfu injection treatment group significantly improved angina pectoris symptoms (OR=3.38, 95%CI： 2.47-4.64, P=0.000) and ischemic ST-T changes in electrocardiogram (OR=3.30, 95%CI： 2.22-4.90, P=0.000), compared with control group. In the Meta-regression analysis, the average age of patients was positively correlated with the improved clinical (ß=0.17) and electrocardiogram (ß=1.15) efficacies. Major complication rate of Shenfu injection was 3.4%, and no serious adverse events were reported. Current clinical evidence in this study proved that Shenfu injection could significantly improve clinical symptoms and ECG ischemic changes for angina pectoris patients, with a good safety.

Angiogenesis Inhibitor, Endostar, Prevents Vasa Vasorum Neovascularization in a Swine Atherosclerosis Model.

AIM: Vasa vasorum neovascularization is a key feature of atherosclerosis (AS) and is strongly associated with inflammatory infiltration, lipid deposition, intraplaque hemorrhage, and hemosiderin deposit. Here we investigate the effects of Endostar, a strong anti-angiogenic drug, on vasa vasorum neovascularization in the experimental porcine model of early AS. METHODS: Eighteen adult male Ba-Ma mini pigs were randomized into three groups, with six animals in each group. The pigs in the normal (N) group were fed a normal diet for 18 weeks, without balloon injury surgery. The animals in the atherosclerotic (AS) control and AS＋Endostar groups were fed a hypercholesterolemic diet for 12 weeks after balloon injury surgery; they received either saline or Endostar for an additional six weeks, while continuing the hypercholesterolemic diet. The atherosclerotic abdominal aorta and levels of serum lipids, TNF-alpha, IL-6, and hs-CRP were analyzed at 18 weeks. RESULTS: The AS group had a significantly higher body weight and serum lipid concentration levels than the N group (p ＜ 0.05), confirming the success of the hypercholesterolemic diet. However, no statistical differences were noted between the AS and AS＋Endostar groups. Histopathology results revealed that vasa vasorum density and intima-media thickness (IMT) had also increased in the AS group compared with those in the N group (p ＜ 0.05). The Endostar treatment significantly alleviated AS with decreased vasa vasorum density and IMT (AS vs. AS＋Endostar, p ＜ 0.05). Western blot analysis indicated that the expression of VEGF, ß-catenin, and TNF-alpha in the atherosclerotic abdominal aorta was considerably reduced by the Endostar treatment. In addition, immunohistochemistry results showed that the angiogenesis markers VEGF and ß-catenin were predominately localized in endothelial cells of the adventitial vasa vasorum. The levels of the serum inflammatory markers TNF-alpha, hs-CRP, and IL-6 were markedly higher in the AS group than in the N group (p ＜ 0.05) but showed no marked difference during the Endostar treatment, suggesting that the local inhibition of angiogenesis was not accompanied by a change in serum inflammatory markers and that the inhibitive effect of Endostar on local TNF-alpha expression could be because of the prevention of vasa vasorum neovascularization. CONCLUSIONS: Our results demonstrated that the Endostar treatment inhibited vasa vasorum neovascularization and AS progression in the experimental porcine model of early AS, supporting the role of vasa vasorum neovascularization in the development of AS and the therapeutic potential of anti-angiogenesis intervention in AS.

Clinicopathologic analysis of POEMS syndrome and related diseases.

BACKGROUND: POEMS syndrome, a rare paraneoplastic disease, is related to multiple organs, multiple systems, and multiple disciplines and can be mistaken for other disorders. Consequently, the diagnoses are often delayed. In this work we studied the clinicopathologic characteristics of the POEMS syndrome to improve early diagnosis to prevent irreversible damage. PATIENTS AND METHODS: We conducted a clinicopathologic analysis of 9 cases of POEMS and made a differential diagnosis with related diseases. RESULTS: The patients with POEMS syndrome were shown to have complicated clinical characteristics, including peripheral neuropathy, organomegaly, endocrinopathy, monoclonal plasma proliferative disorder, skin changes, extravascular volume overload, sclerotic bone lesions, thrombocytosis, and Castleman disease. POEMS syndrome shared many elements with other diseases and the key way to differentiate them was to determine whether there were other fundamental POEMS syndrome symptoms or signs. The level of M-protein in serum and plasma cells in bone marrow of POEMS patients was lower than that of patients with multiple myeloma (MM). Sclerotic bone lesions were a distinctive feature in patients with POEMS, compared with in those with MM. CONCLUSION: Some unique clinicopathologic characteristics of POEMS syndrome can be used for differential diagnosis. This study provides increased awareness of POEMS syndrome.

Right ventricular apical obstruction in a patient with hypereosinophilia: Löffler endocarditis.

Idiopathic hypereosinophilic syndrome (HES) is a rare leukoproliferative systemic disorder characterized by sustained overproduction of eosinophils and poor prognosis. A case that a 67-year-old man with persistent symptoms of heart failure due to cardiac involvement in idiopathic HES is concentrated on. Echocardiography revealed the marked endocardial thickening of both ventricles with an apical obstruction of the right ventricle. Medical therapy, including low dose dopamine and furosemidum, was initiated with corticosteroids, imatinib and hydroxycarbamide. Remission of symptoms had persisted for only 3 weeks. As the count of eosinophils rebounded, the patient suffered with refractory heart failure, severe hypoxemia and acute renal insufficiency, eventually died 62 days after his hospitalization. The rechecking of his last MRI showed thrombus both in right atrium and superior vena cava, which indicated that he might have died of pulmonary embolism, besides the refractory heart failure and multiple organ failure.

Endostar, a modified recombinant human endostatin, suppresses angiogenesis through inhibition of Wnt/ß-catenin signaling pathway.

Endostar, a novel modified recombinant human endostatin, is now widely studied for the treatment of diseases that are characterized or caused by pathological angiogenesis. However, its molecular mechanism remains unclear. In this study, we investigated the effects of Endostar on the Wnt/ß-catenin signaling pathway, which has emerged as an important aspect of angiogenesis. We showed that Endostar significantly inhibited the proliferation, migration, invasion, and capillary-like tube formation of human umbilical vascular endothelial cells in a dose-dependent manner. Using a luciferase reporter assay, we also demonstrated that Endostar suppressed ß-catenin-dependent T cell factor transcriptional activity in increasing doses. Moreover, we found that Endostar treatment also restricted the stabilized mutant ß-catenin-mediated increase in transcriptional activity, suggesting that Endostar exerts its inhibitory influence on Wnt/ß-catenin signaling by targeting ß-catenin or its downstream molecules. Western blot and immunofluorescence results revealed that Endostar significantly decreased nuclear and total ß-catenin levels. Finally, we discovered that Endostar down-regulated cyclin D1 and VEGF, two proteins that are known as the downstream targets of Wnt/ß-catenin signaling and that also play important roles in angiogenesis. Our findings suggested that Endostar inhibits angiogenesis and that the downregulation of the Wnt/ß-catenin signaling pathway may be involved in the inhibition of angiogenesis by Endostar. These results support the use of Endostar in further clinical applications.

Inhibitory effect of cryptotanshinone on angiogenesis and Wnt/ß-catenin signaling pathway in human umbilical vein endothelial cells.

OBJECTIVE: To investigate the anti-angiogenic effect of cryptotanshinone (CPT) on human umbilical vein endothelial cells (HUVECs) and the effect of CPT on Wnt/ß-catenin signaling pathway. METHODS: HUVECs were incubated with 0, 2.5, 5, 10, and 20 µ mol/L CPT for detecting cell viability with dimethyl thiazolyl-2,5-diphenyltetrazolium bromide (MTT) assay. Then, HUVECs were incubated with 0, 2.5, 5, and 10 µ mol/L CPT for detecting endothelial cell migration, invasion, and tubular-like structure formation with wound healing, transwell invasion and matrigel tube formation assays, respectively. To gain insight into CPT-mediated signaling, the effects of CPT on T-cell factor/lymphocyte enhancer factor (TCF/LEF) transcription factors were detected by the Dual-luciferase reporter assay. Next, the nuclear expression of ß-catenin was evaluated using Western blot and immunochemistry. Finally, vascular endothelial growth factor (VEGF) and cyclin D1, downstream proteins of the Wnt pathway were examined with Western blot. RESULTS: CPT dose-dependently suppressed endothelial cell viability, migration, invasion, and tubular-like structure formation. In particular, CPT blocked ß-catenindependent transcription in HUVECs in a dose-dependent manner. In Western bolt, 10 µ mol/L CPT decreased expression of ß-catenin in nucleus of HUVECs (P<0.01). In immunohistochemistry, ß-catenin was more potent in response to LiCl (an activator of the pathway) treatment. However, the signals were weaker in the nucleus of the CPT (10 µ mol/L) group, compared to the positive control. Also, VEGF and cyclin D1 were both eliminated by CPT in 5 and 10 µ mol/L doses (P<0.05). CONCLUSION: Our study supported the role of CPT as an angiogenic inhibitor, which may impact on the Wnt/ß-catenin signaling pathway.