VEGF-B prevents excessive angiogenesis by inhibiting FGF2/FGFR1 pathway.

Although VEGF-B was discovered as a VEGF-A homolog a long time ago, the angiogenic effect of VEGF-B remains poorly understood with limited and diverse findings from different groups. Notwithstanding, drugs that inhibit VEGF-B together with other VEGF family members are being used to treat patients with various neovascular diseases. It is therefore critical to have a better understanding of the angiogenic effect of VEGF-B and the underlying mechanisms. Using comprehensive in vitro and in vivo methods and models, we reveal here for the first time an unexpected and surprising function of VEGF-B as an endogenous inhibitor of angiogenesis by inhibiting the FGF2/FGFR1 pathway when the latter is abundantly expressed. Mechanistically, we unveil that VEGF-B binds to FGFR1, induces FGFR1/VEGFR1 complex formation, and suppresses FGF2-induced Erk activation, and inhibits FGF2-driven angiogenesis and tumor growth. Our work uncovers a previously unrecognized novel function of VEGF-B in tethering the FGF2/FGFR1 pathway. Given the anti-angiogenic nature of VEGF-B under conditions of high FGF2/FGFR1 levels, caution is warranted when modulating VEGF-B activity to treat neovascular diseases.

An Efficient UPLC-MS/MS Method Established to Detect Relugolix Concentration in Rat Plasma.

Relugolix, a gonadotropin-releasing hormone (GnRH) receptor antagonist, has been well studied in the treatment of endometriosis symptomatic. It is mainly metabolized by the CYP3A subfamily of P450 enzymes, while minorly metabolized by CYP2C8. Daidzein in different dose groups exhibited a certain induction on the mRNA expression level of CYP3A4 and resulted in the potent induction of CYP3A4. However, it is still unknown whether daidzein and relugolix interact. We developed an effective ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method to study the effect of daidzein on the pharmacokinetics of relugolix in rats after oral administration of 12 mg/kg relugolix in a single or mixed of 50 mg/kg daidzein. The results showed that the method had respectable linearity (r 2 > 0.999) on the scale of 0.7-1000 ng/mL. The intra-day precision was between 3.0% and 8.4% in this assay, and the inter-day was between 4.0% and 11.7%. The intra-day accuracy was from -4.3% to 6.1%, and the inter-day was 2.9% to 12.1%. Another three key indicators, including the stability, the recovery rate of extraction and the new technique's matrix effect, were perfectly in accord with the test verification rule in the biological medium by the United States Food and Drug Administration. Meanwhile, treatment with daidzein led to a decrease in Cmax and AUC0-t of relugolix by about 15.56% and 21.36%, respectively. Although there was no statistical difference in pharmacokinetic parameters, it reflected the induction trend of daidzein on relugolix metabolism for food-drug interaction. It would provide reference and improvement value for subsequent experiments.

Incremental significance and sex discrepancies of neck circumference on the odds of ischaemic stroke: a multistage, population-based, cross-sectional study from Northeast China.

OBJECTIVES: Accumulated evidence suggests that neck circumference (NC) is associated with cardiometabolic risk factors. However, limited studies are available regarding the association between NC or height normalised NC (neck-to-height ratio (NHR)) and risk of ischaemic stroke (IS) in the Chinese population. Therefore, we aimed at examining the associations between NC or NHR and odds of IS and exploring the discrepancies between men and women. DESIGN: A multistage cluster cross-sectional study. SETTING: A population-based study carried out in Northeast China. METHODS: A cross-sectional study was undertaken in Northeast China between September 2017 and March 2019, involving 7236 men and 11 352 women, respectively. The median age of participants was 60.30 years, ranging from 40 to 97 years. The associations between NC or NHR and odds of IS were calculated using multiple logistic regression models. Dose-response relationships were depicted using restricted cubic spline functions. Reclassification analyses were carried out to determine the incremental significance of NC or NHR on the odds of IS. RESULTS: In women, NC and NHR were significantly associated with the odds of IS, independent of traditional risk factors and other anthropometric parameters for obesity. The highest quartile of NC and NHR had a 1.60 (95% CI 1.16 to 2.22)-and 1.72 (95% CI 1.23 to 2.41) times higher odds of IS compared with the lowest quartile. Furthermore, the odds of IS increased by 1.10 (95% CI 1.01 to 1.20) and 1.12 (95% CI 1.02 to 1.22) times per 1 SD increase in NC and NHR, respectively. Reclassification analyses showed that the proportion of correct classification increased by 11.5% (95% CI 2.2% to 20.7%) and 22.8% (95% CI 13.5% to 32.0%) after the addition of NC or NHR into established models, respectively. However, the findings could not be replicated in men. CONCLUSION: NC and NHR might be promising independent indicators for women IS. Their incremental value in the risk stratification of IS enables the individualised prevention of IS in women.

Sex-specific prediction value of common carotid artery diameter for stroke risk in a hypertensive population: a cross-sectional study.

BACKGROUND: The importance of sex as a risk factor for stroke has been established. This study aimed to assess sex-related disparities in carotid artery diameter and stroke in a hypertensive population. METHODS: The cross-sectional survey was conducted in rural areas of northeast China. A multistage cluster sampling method was employed to select a representative population. The study comprised 3,245 individuals with hypertension. The common carotid artery (CCA) interadventitial diameter was measured by ultrasound. A linear model of restricted cubic spline function was used to characterize the concentration-response (C-R) relationship between CCA diameter and stroke. RESULTS: The overall prevalence of stroke was 8.9% among hypertensive individuals, with a higher rate in men than in women (10.8% vs. 7.6%). When the women's CCA diameters were divided into quartiles, the top quartile (>8.10 mm) had a 2.49 (95% CI: 1.36-4.56) times greater risk of stroke compared to the bottom quartile (≤6.80 mm) after adjustment was made for other variables. The C-R relationship further confirmed a positive association between CCA diameter and stroke prevalence in women. Moreover, a category-free net reclassification index (0.325; 95% CI: 0.173-0.476; P<0.001) and an integrated discrimination index (0.008; 95% CI: 0.004-0.012, P<0.001) showed improvement in predicting the probability of stroke from CCA diameter. However, no significant relationship between CCA diameter and prevalence of stroke was found in men. CONCLUSIONS: The risk of stroke increased proportionally with the enlargement of the CCA diameter in women, supporting the sex-specific value of CCA diameter in optimizing the risk stratification of stroke.

High Burden of Carotid Atherosclerosis in Rural Northeast China: A Population-Based Study.

Objective: Carotid atherosclerosis is a known marker of increased cardiovascular risk. We aimed to assess the current epidemiology of carotid atherosclerosis, carotid plaque and related risk factors in rural northeast China. Methods: The population-based, cross-sectional study was conducted in 5,838 adults aged ≥40 years residing in rural northeast China in 2017-2018. A multi-stage cluster sampling method was used to select the representative sample. Carotid atherosclerosis was defined as carotid intima-media thickness (CIMT) ≥1.0 mm or presence of plaque. Results: The mean CIMT was 0.72 ± 0.13 mm and increased with age in this population. Among 2,457 individuals with carotid atherosclerosis, 2,333 were diagnosed with carotid plaque, and 210 individuals were moderate or severe carotid stenosis. Crude prevalence of carotid atherosclerosis and plaque were 42.1 and 40.0%, significantly higher in men than in women (p < 0.001). The age-standardized prevalence of carotid atherosclerosis and carotid plaque were 33.1 and 31.5%, respectively. Advancing age, men, hypertension, diabetes, current smoking, ever-smoking and lack of exercise were risk factors for carotid atherosclerosis. Hypertension (69.1%), dyslipidemia (26.0%) and diabetes (16.1%) were highly prevalent in participants with carotid atherosclerosis. However, the control rates of those comorbidities were frustratingly low (4.7, 8.2, and 14.2%, respectively). Conclusions: The high prevalence of carotid atherosclerosis, carotid plaque, carotid stenosis and uncontrolled risk factors indicated the high burden of cardiovascular disease in rural northeast China, particularly in men. Strategies of prevention and management of atherosclerosis and related risk factors were urgently needed in rural northeast China.

Estimate of prevalent ischemic stroke from triglyceride glucose-body mass index in the general population.

BACKGROUND: To investigate the relationship between triglyceride glucose-body mass index (TyG-BMI) and ischemic stroke. METHODS: Leveraging two Chinese general population surveys, the Northeast China Rural Cardiovascular Health Study (NCRCHS, N = 11,097) and the National Stroke Screening and Intervention Program in Liaoning (NSSIPL, N = 10,862), we evaluated the relationship between TyG-BMI and ischemic stroke by a restricted cubic spline and multivariate logistic regression after adjusting age, sex, level of education, exercise regularly, current smoking, current drinking, atrial fibrillation, hypertension, coronary artery disease, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol. The category-free analysis was used to determine whether TyG-BMI enhanced the capacity of estimating ischemic stroke. RESULTS: A total of 596 and 347 subjects, respectively, from NSSIPL and NCRCHS were survivors of ischemic stroke. In NSSIPL, the relationship between TyG-BMI and ischemic stroke was linear and did not have a threshold or saturation effect according to the results of the restricted cubic spline. The regression analysis indicated that the risk of ischemic stroke increased 20% for per SD increase of TyG-BMI after multivariate adjustment [odds ratio (OR): 1.20, 95% confidence interval (CI): 1.10-1.32]. Compared with those in the lowest tertile, the risk of ischemic stroke in subjects with intermediate and high TyG-BMI was significantly higher [OR (95% CI): 1.39 (1.10-1.74); OR (95% CI) 1.72 (1.37-2.17), respectively]. Category-free analysis indicated that TyG-BMI had a remarkable improvement in the ability to estimate prevalent ischemic stroke [NRI (95% CI): 0.188 (0.105-0.270)]. These abovementioned relationships were confirmed in NCRCHS. CONCLUSIONS: The present study found the robust correlation between TyG-BMI and ischemic stroke, independently of a host of conventional risk factors. Meanwhile, our findings also suggested the potential usefulness of TyG-BMI to improve the risk stratification of ischemic stroke.

Prevalence and determinants of metabolic syndrome based on three definitions in rural northeast China.

OBJECTIVE: To gain more comprehensive understanding of metabolic syndrome (Mets) among in general Chinese population. DESIGN: Cross-sectional study. Mets was defined by three widely accepted definitions including modified Adults Treatment Panel (ATP) III criteria, International Diabetes Federation (IDF) criteria and harmonized definition. Risk factors were evaluated by using multivariate logistic regression. SETTING: Nineteen rural villages in northeast China. PARTICIPANTS: The survey was conducted in September 2017 and May 2018 on 10 926 individuals. RESULTS: According to modified ATP III criteria, IDF criteria and harmonised definition, the overall prevalence of Mets was 41·3 % (95 % CI 40·3, 42·2), 34·2 % (95 % CI 33·2, 35·1) and 44·1 % (95 % CI 43·1, 45·1), respectively. Females had a higher prevalence, and elevated blood pressure was the most frequent. Age, female sex, non-peasant worker, higher BMI and lower-annual income were independent risk factors of Mets in all three definitions (all ps < 0·05). Based on modified ATP III criteria and harmonised definition, heavy drinking was positively correlated with Mets. In contrast, former drinking was inversely associated with Mets. CONCLUSIONS: Mets is highly prevalent in rural areas of northeast China. Its independent risk factors include higher age, female sex, non-peasantry worker, higher BMI and lower-annual income. Modified ATP III criteria and harmonised definition may be superior definitions of Mets.

Isolation of T cell receptor specifically reactive with autologous tumour cells from tumour-infiltrating lymphocytes and construction of T cell receptor engineered T cells for esophageal squamous cell carcinoma.

BACKGROUND: T cell receptor-engineered T cells (TCR-Ts) therapy is a promising cancer treatment strategy. Nowadays, most studies focused on identification of high-avidity T cell receptors (TCRs) directed against neoantigens derived from somatic mutations. However, few neoantigens per patient could induce immune response in epithelial cancer and additionally many tumor-specific antigens could be derived from noncoding region. Autologous tumor cells (ATCs) could be unbiased stimulators in activating and enriching tumor-reactive T cells. However, it's unknown if T cells engineered to express TCRs isolated from tumor-reactive T cells enriched by ATCs have strong antitumor response. METHODS: In this study, multiple TIL fragments obtained from a patient with esophageal squamous cell carcinoma (ESCC) were screened for specific recognition of ATCs. Tumor-reactive TILs were enriched by in vitro repeated stimulation of ATCs and isolated based on CD137 upregulation. Subsequently, tumor-reactive TCR was obtained by single-cell RT-PCR analysis and was introduced into peripheral blood lymphocytes to generate TCR-Ts. RESULTS: We found that phenotype and effect function of TIL fragments derived from different tumor sites were spatially heterogeneous. Of four TIL fragments, only TIL-F1 could specifically identify ATCs. Subsequently, we isolated CD8+ CD137+ T cells from pre- and post-stimulated TIL-F1 co-cultured with ATCs, and identified their most dominant TCR. This TCR was introduced into PBLs to generate TCR-Ts, which specifically identified and killed ATCs in vivo and in vitro. CONCLUSION: This strategy provides the means to generate tumor-reactive TCR-Ts for ESCC, which is especially important for patients without prior knowledge of specific epitopes and might be applied for other cancers.

Prevalence of Obesity and Associated Risk Factors and Cardiometabolic Comorbidities in Rural Northeast China.

OBJECTIVE: To investigate the epidemiological features of obesity in rural Northeast China. DESIGN: This was a 2017-2018 cross-sectional study of 10,891 participants aged ≥40 years that was designed to investigate the prevalence of obesity in rural areas of Liaoning Province. Demographic data, biochemical parameters, and physical examinations were completed by well-trained personnel. Logistic regression analyses were then carried out to investigate independent risk factors and associated cardiometabolic comorbidities of obesity. RESULTS: The proportions of general obesity only, central obesity only, and combined obesity were 1.0%, 31.3%, and 17.4%, respectively. Overall, 49.8% of our subjects were obese. Female gender, being married, being separated/divorced/widowed, or eating more meat were significantly associated with obesity. Smoking, higher family income, or regular physical exercise were negatively associated with obesity. General obesity only was significantly correlated with hypertension, diabetes, and high triglycerides (OR = 2.79, OR = 2.79, and OR = 3.37, resp.). General obesity only was irrelevant to high total cholesterol, low high-density lipoprotein cholesterol, and high low-density lipoprotein cholesterol, although central obesity only, or combined obesity, was relevant to these factors. Prehypertension and prediabetes showed a positive association with different types of obesity. CONCLUSIONS: We identified a high prevalence of general and central obesity in rural Northeast China, with similar independent risk factors. Participants with combined obesity had the highest risk of cardiometabolic comorbidities, indicating that the combined use of both waist circumference and body mass index is useful in practice.

Low Infrared Emissivity Coating Based on Graphene Surface-Modified Flaky Aluminum.

A low infrared emissivity coating was prepared using graphene surface-modified flaky aluminum complex powders (rGO-FAl) as fillers. The flaky aluminum powders were coated with graphene through chemical bonding. Compared with pure flaky aluminum, the Vis-NIR diffuse reflectance of rGO-FAl complex powders was significantly decreased, which was beneficial to the low glossiness of the coating. After the modification, the glossiness at 60° of the coating with 40% (mass fraction) pigments decreased from 12.8 to 6.7, while the coating maintained low infrared emissivity (0.238~0.247) at a spectral range of 8⁻14 µm. In the electrochemical impedance spectroscopy (EIS) measurement, at the lowest frequency, the impedance of the Al-rGO test plate was at least two orders of magnitude greater than that of the control Al test plate, and the graphene layer significantly increased the bandwidth of the maximum phase angle, which indicates a good protective effect of the ultra-thin graphene layer on metal in a corrosive environment. The coating with 40% rGO-FAl complex powders can maintain its appearance after 500 h of salt spray corrosion testing. In contrast, the color of the coating with the original aluminum powders changed after only 300 h.

Preparation of reduced graphene oxide coated flaky carbonyl iron composites and their excellent microwave absorption properties.

In this study, a novel absorber with a high performance microwave absorption property was prepared by innovatively coupling flaky carbonyl iron (FCI) and reduced graphene oxide (rGO) nanosheets into a homogenous composite. The rGO nanosheets are tightly coated on the surface of FCI, which gives typical dielectric dispersion behavior of complex permittivity and resultantly optimizes characteristic impedance matching. Meanwhile, the introduction of rGO as a dielectric lossy material endows FCI with improved dielectric loss ability and unfading magnetic loss ability. In the frequency range of 2.0-18.0 GHz, the rGO-coated FCI composite gives a minimum value of reflection loss with -65.4 dB at 5.2 GHz when the absorber thickness is 3.87 mm and always shows effective bandwidth under -20 dB when absorber thickness is from 1.5 mm to 5 mm. The contribution of typical dielectric dispersion behavior in rGO-coated FCI is demonstrated by a delta-function method to characteristic impedance matching and the excellent microwave absorbing performance.

Critical role of caveolin-1 in ocular neovascularization and multitargeted antiangiogenic effects of cavtratin via JNK.

Ocular neovascularization is a devastating pathology of numerous ocular diseases and is a major cause of blindness. Caveolin-1 (Cav-1) plays important roles in the vascular system. However, little is known regarding its function and mechanisms in ocular neovascularization. Here, using comprehensive model systems and a cell permeable peptide of Cav-1, cavtratin, we show that Cav-1 is a critical player in ocular neovascularization. The genetic deletion of Cav-1 exacerbated and cavtratin administration inhibited choroidal and retinal neovascularization. Importantly, combined administration of cavtratin and anti-VEGF-A inhibited neovascularization more effectively than monotherapy, suggesting the existence of other pathways inhibited by cavtratin in addition to VEGF-A. Indeed, we found that cavtratin suppressed multiple critical components of pathological angiogenesis, including inflammation, permeability, PDGF-B and endothelial nitric oxide synthase expression (eNOS). Mechanistically, we show that cavtratin inhibits CNV and the survival and migration of microglia and macrophages via JNK. Together, our data demonstrate the unique advantages of cavtratin in antiangiogenic therapy to treat neovascular diseases.

PDGF-CC underlies resistance to VEGF-A inhibition and combinatorial targeting of both suppresses pathological angiogenesis more efficiently.

Anti-VEGF-A therapy has proven to be effective for many neovascular diseases. However, drug resistance to anti-VEGF-A treatment can develop. Also, not all patients with neovascular diseases are responsive to anti-VEGF-A treatment. The mechanisms underlying these important issues remain unclear. In this study, using different model systems, we found that inhibition of VEGF-A directly upregulated PDGF-CC and its receptors in multiple cell types in pathological angiogenesis in vitro and in vivo. Importantly, we further revealed that combinatorial targeting of VEGF-A and PDGF-CC suppressed pathological angiogenesis more efficiently than monotherapy. Given the potent angiogenic activity of PDGF-CC, our findings suggest that the development of resistance to anti-VEGF-A treatment may be caused by the compensatory upregulation of PDGF-CC, and combined inhibition of VEGF-A and PDGF-CC may have therapeutic advantages in treating neovascular diseases.

Inactivation of PI3Kδ induces vascular injury and promotes aneurysm development by upregulating the AP-1/MMP-12 pathway in macrophages.

OBJECTIVE: An aneurysm is an inflammatory vascular condition. Phosphatidylinositol 3-kinases δ is highly expressed in leukocytes, and play a key role in innate immunity. However, the link between phosphatidylinositol 3-kinases δ and aneurysm development has not yet been elucidated. APPROACH AND RESULTS: Carotid ligation unexpectedly induced characteristic aneurysm formation beneath the ligation point in p110δ(D910A/D910A) mice (n=25; P<0.001 versus wild-type). Besides, p110δ inactivation exacerbated CaCl2-induced abdominal aortic aneurysms development. A reverse transcription polymerase chain reaction microarray revealed significant extracellular matrix components degradation and matrix metalloproteinases (MMPs) upregulation in the abdominal aorta of p110δ(D910A/D910A) mice. Similarly, the expression of both collagen I and IV was significantly decreased (n=10; P<0.05 versus wild-type) in carotid artery. Western blot assay confirmed that MMP-12 was significantly upregulated in arteries of p110δ(D910A/D910A) mice (n=10; P<0.01 versus wild-type). In vitro, p110δ inactivation marked increase peritoneal macrophages recruitment and synergistically enhance tumor necrosis factor-α-induced recruitment. A specific phosphatidylinositol 3-kinases δ inhibitor (IC87114) or genetic p110δ inactivation upregulated MMP-12 expression and c-Jun phosphorylation (n=6; P<0.05 versus wild-type macrophages). IC87114 also increased activator protein-1 DNA-binding activity (n=6; P<0.001 versus control) and enhanced the effect of tumor necrosis factor-α on activator protein-1-binding activity (n=5; P<0.01 versus tumor necrosis factor-α treatment groups). Knockdown of c-Jun suppressed the effect of the IC87114 and tumor necrosis factor-α on MMP-12 mRNA expression (n=5 in each group; P<0.01 versus scrRNA treatment groups). CONCLUSIONS: Our findings demonstrate that p110δ inactivation leads to extracellular matrix degradation in vessels and promotes aneurysm development by inducing macrophages migration and upregulating the activator protein-1/MMP-12 pathway in macrophages.