Acteoside isolated from Colebrookea oppositifolia attenuates I/R brain injury in Wistar rats via modulation of HIF-1α, NF-κB, and VEGF pathways.

AIMS: The objective of this study was to assess the anti-stroke activity of acteoside isolated from methanolic root extract of C. oppositifolia METHODS: Ischemia-reperfusion(I/R) brain injury was induced in Wistar rats to assess the anti-stroke activity of acteoside. Rats were pretreated with acteoside (10, 25 & 50 mg/kg, p.o.) before the induction of I/R injury. Parameters such as neurological, motor-cognitive functions were evaluated along with morphological (brain volume, infarct size), biochemical (SOD, Catalase, GSH, lipid peroxidation, TNF-α, IL-6, IL-10, ICAM-1, HIF-1α, VEGF, and NF-κB), histopathological, and gene expression studies (HIF-1α, VEGF) were performed to study the protective effect of acteoside against I/R induced brain injury. RESULTS: I/R injury caused significant deterioration of neurological (p < 0.01), motor (p < 0.01) and cognitive (p < 0.01) functions, associated with increase in the brain volume (p < 0.01), and infarct size (p < 0.01); increase in the levels of MDA, TNF-α, IL-6, ICAM-1, HIF-1α, VEGF, and NF-κB along with significant decrease in SOD, catalase, GSH, and IL-10 (p < 0.01 for all parameters) compared to Sham control group. Histology of brain tissue of disease control group exhibited significant vascular changes, neutrophil infiltration, cerebral oedema, and necrosis of the neuronal cells. Further, the gene-expression studies showed significant increase in the HIF-1α (p < 0.01) and VEGF (p < 0.01) mRNA levels in the I/R control compared to Sham control. Interestingly, the acteoside (10, 25 & 50 mg/kg) has prevented the neurological, motor and cognitive dysfunctions, along with inhibiting the morphological, biochemical, histological and gene expression changes induced by I/R-injury (p < 0.05 for 10 mg; p < 0.01 for 25 & 50 mg/kg of acteoside for all the parameters). CONCLUSION: These findings suggest that acteoside possess potent anti-stroke activity through modulation of HIF-1α, NF-κB, and VEGF pathway along with its potent antioxidant activity.

Carbon ion radiotherapy of human lung cancer attenuates HIF-1 signaling and acts with considerably enhanced therapeutic efficiency.

Carbon ion irradiation is an emerging therapeutic option for various tumor entities. Radiation resistance of solid tumors toward photon irradiation is caused by attenuation of DNA damage in less oxygenated tumor areas and by increased hypoxia-inducible factor (HIF)-1 signaling. Carbon ion irradiation acts independently of oxygen; however, the role of HIF-1 is unclear. We analyzed the effect of HIF-1 signaling after carbon ions in comparison to photons by using biological equivalent radiation doses in a human non-small-cell cancer model. The studies were performed in cultured A549 and H1299 cell lines and in A549 xenografts. Knockdown of HIF-1α in vivo combined with photon irradiation delayed tumor growth (23 vs. 13 d; P<0.05). Photon irradiation induced HIF-1α and target genes, predominantly in oxygenated cells (1.6-fold; P<0.05), with subsequent enhanced tumor angiogenesis (1.7-fold; P<0.05). These effects were not observed after carbon ion irradiation. Micro-DNA array analysis indicated that photons, but not carbon ions, significantly induced components of the mTOR (mammalian target of rapamycin) pathway (gene set enrichment analysis; P<0.01) as relevant for HIF-1α induction. After carbon ion irradiation in vivo, we observed substantially decreased HIF-1α levels (8.9-fold; P<0.01) and drastically delayed tumor growth (P<0.01), an important finding that indicates a higher relative biological effectiveness (RBE) than anticipated from the cell survival data. Taken together, the evidence showed that carbon ions mediate an improved therapeutic effectiveness without tumor-promoting HIF-1 signaling.

The relationship between renal function and surgical outcomes of patients with proliferative diabetic retinopathy.

Objectives: The relationship between renal function and diabetic retinopathy has been controversial. This study is to investigate the influence of renal function on the complex and surgical outcomes of proliferative diabetic retinopathy (PDR). Methods: This was a post hoc analysis of the CONCEPT clinical trial. A total of 45 eyes with PDR underwent vitrectomy were included. Based on the estimated glomerular filtration rate (eGFR), they were divided into abnormal renal function group (ARF group) and normal renal function group (NRG group). Baseline PDR complex, intraoperative outcomes (Intraoperative bleeding, frequency of endodiathermy, surgical time, iatrogenic hole, and tamponade) and postoperative outcomes (logMAR best-corrected visual acuity, vitreous re-hemorrhage, and macular edema, follow up at postoperative 1 month and 3 months) were estimated. Vitreous, aqueous humor and serum were collected at the vitrectomy day and Vascular endothelia growth factor-A levels were quantified for all included patients using liquid chip method. Results: There was no significant difference in baseline PDR complex, intraoperative and postoperative outcomes between ARF group and NRG group (all P > 0.05). At the vitrectomy day, there was also no difference of Vascular endothelia growth factor-A levels in vitreous, aqueous humor and serum between the two groups (all P > 0.05). Conclusion: Our results showed that the renal function seems not parallel to the severity of PDR, neither to the surgical outcomes. This might be interpreted by the similar Vascular endothelia growth factor-A levels in vitreous, aqueous humor and serum between the two groups.

OxLDL promotes lymphangiogenesis and lymphatic metastasis in gastric cancer by upregulating VEGF­C expression and secretion.

Gastric cancer is one of the most malignant tumor types, and its metastasis is a notable cause of mortality. Among the methods of tumor metastasis, lymphatic metastasis is the predominant one in gastric cancer. A previous study reported that the plasma oxidized low­density lipoprotein (oxLDL) is the risk factor associated with the development of tumors in patients with abnormal lipid metabolism, but the influence of plasma oxLDL in the lymphatic metastasis of gastric cancer remains unclear. In the present study, the concentration of plasma oxLDL from patients with gastric cancer was detected with an ELISA kit, and the lymphatic vessel density in gastric cancer tissues was determined by D2­40 staining. The correlation analysis of oxLDL concentration and lymphatic vessel density demonstrated that plasma oxLDL was positively correlated with lymphatic metastasis in patients with gastric cancer. Subsequently, the popliteal lymph node metastasis animal experiment with nude mice confirmed that oxLDL could promote the lymphatic metastasis of gastric cancer. Following this, the western blotting and ELISA data demonstrated that oxLDL promoted the expression and secretion of vascular endothelia growth factor (VEGF)­C in gastric cancer cell lines. Finally, blocking the lectin­like oxLDL­1 (LOX­1) receptor, a specific receptor for oxLDL, and the nuclear factor (NF)­κB signaling pathway following oxLDL (50 µg/ml) treatment in HGC­27 cells revealed that oxLDL could activate the NF­κB signaling pathway mediated by LOX­1, with subsequent upregulation of VEGF­C expression, and secretion in and from gastric cancer cells, and finally that it could promote the lymphatic metastasis of gastric cancer. These data indicate the association between the plasma oxLDL and the lymphatic metastasis of gastric cancer, and indicate that oxLDL elimination may be a potential therapeutic target for the prevention and intervention of early lymph node metastasis in gastric cancer.

Autologous chondrocyte grafting promotes bone formation in the posterolateral spine.

BACKGROUND CONTEXT: Pseudarthrosis following spinal fusion remains problematic despite modern surgical and grafting techniques. In surgical spinal fusion, new bone forms via intramembranous and endochondral ossification, with endochondral ossification occurring in the hypoxic zones of the fusion bed. During bone development and fracture healing, the key cellular mediator of endochondral ossification is the hypertrophic chondrocyte given its ability to function in hypoxia and induce neovascularization and ossification. We therefore hypothesize that hypertrophic chondrocytes may be an effective bone graft alternative. PURPOSE: Spinal fusion procedures have increased substantially; yet 5% to 35% of all spinal fusions may result in pseudoarthrosis. Pseudoarthrosis may occur because of implant failure, infection, or biological failure, among other reasons. Advances in surgical techniques and bone grafting have improved fusion; however pseudarthrosis rates remain unacceptably high. Thus, the goal of this study is to investigate hypertrophic chondrocytes as a potential biological graft alternative. METHODS: Using a validated murine fracture model, hypertrophic chondrocytes were harvested from fracture calluses and transplanted into the posterolateral spines of identical mice. New bone formation was assessed by X-ray, microcomputed tomography (µCT), and in vivo fluorescent imaging. Results were compared against a standard iliac crest bone graft and a sham surgery control group. Funding for this work was provided by the Department of Orthopaedics and Rehabilitation, the OREF (Grant #16-150), and The Caitlin Lovejoy Fund. RESULTS: Radiography, µCT, and in vivo fluorescent imaging demonstrated that hypertrophic chondrocytes promoted bone formation at rates equivalent to iliac crest autograft. Additionally, µCT analysis demonstrated similar fusion rates in a subset of mice from the iliac crest and hypertrophic chondrocyte groups. CONCLUSIONS: This proof-of-concept study indicates that hypertrophic chondrocytes can promote bone formation comparable to iliac crest bone graft. These findings provide the foundation for future studies to investigate the potential therapeutic use of hypertrophic chondrocytes in spinal fusion.

Antileukemic effects of midostaurin in acute myeloid leukemia - the possible importance of multikinase inhibition in leukemic as well as nonleukemic stromal cells.

INTRODUCTION: Midostaurin is a multikinase inhibitor that inhibits receptor tyrosine kinases (Flt3, CD117/c-kit, platelet-derived growth factor receptor, vascular endothelial growth factor receptor 2) as well as non-receptor tyrosine kinases (Frg, Src, Syk, Protein kinase C). Combination of midostaurin with conventional intensive chemotherapy followed by one year maintenance monotherapy was recently reported to improve the survival of acute myeloid leukemia (AML) patients with Flt3 mutations. Areas covered: Relevant publications were identified through literature searches in the PubMed database. We searched for (i) original articles describing the results from clinical studies; (ii) published articles describing the importance of midostaurin-inhibited kinases for leukemogenesis and chemosensitivity. Expert opinion: Midostaurin monotherapy is well tolerated, combined with conventional chemotherapy gastrointestinal toxicity increases significantly. Midostaurin alters anthracycline pharmacokinetics. Furthermore, its antileukemic effects may not only be mediated through Flt3 inhibition alone; the inhibition of other kinases may also be important for the overall antileukemic effect. Midostaurin may then have direct effects on the leukemic cells but also indirect antileukemic effects through inhibition of the AML-supporting effects of neighboring stromal cells in the bone marrow microenvironment. Midostaurin may thus be used in combination with intensive chemotherapy, as maintenance treatment or as disease-stabilizing treatment for elderly unfit patients.

Comparison on serum biomarkers for anovulatory and ovulatory dysfunctional uterine bleeding in Lizu females.

OBJECTIVE: To screen, identify, and compare the serum biomarkers between anovulatory dysfunctional uterine bleeding (ADUB) and ovulatory dysfunctional uterine bleeding (ODUB) in Lizu females. METHODS: The subjects included 128 ADUB patients, 63 ODUB patients, and 93 controls. The serum and supernate of the subjects' mense were collected and stored at -80 °C until use. Differential proteins in the sera of three groups were screened using surface-enhanced laser desorption ionization time-of-flight mass spectrometry. The screened proteins were then identified by tricine-SDS-PAGE gel and spectrometry. Protein expression levels in the menses of ADUB, ODUB, and control subjects were determined using ELISA, RT-PCR, and Western blotting. SPSS 14.1 was used for statistical analysis and chart drawing (α = 0.05). RESULTS: Three differential protein peaks with peak values of 11.80, 13.59, and 14.68 km/z were screened and identified as serum amyploid protein A (SAA), vascular endothelial growth factor, and vitamin K epoxide reductase, respectively. The SAA was highly expressed in the menses of ADUB and ODUB patients but poorly expressed in the controls. The vascular endothelial growth factor was highly expressed in the menses of ODUB and controls but poorly expressed in ADUB patients. Meanwhile, the vitamin K epoxide reductase was highly expressed in the menses of ADUB and control subjects but poorly expressed in ODUB patients. CONCLUSIONS: The SAA is the common serum biomarker of ADUB and ODUB. ADUB may be related to angiogenesis impairment, whereas ODUB may be associated with blood coagulation disruption.

Gene and protein therapies utilizing VEGF for ALS.

Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease that is usually fatal within 2-5years. Unfortunately, the only treatment currently available is riluzole, which has a limited efficacy. As a redress, there is an expanding literature focusing on other potential treatments. One such potential treatment option utilizes the vascular endothelial growth factor (VEGF) family, which includes factors that are primarily associated with angiogenesis but are now increasingly recognized to have neurotrophic effects. Reduced expression of a member of this family, VEGF-A, in mice results in neurodegeneration similar to that of ALS, while treatment of animal models of ALS with either VEGF-A gene therapy or VEGF-A protein has yielded positive therapeutic outcomes. These basic research findings raise the potential for a VEGF therapy to be translated to the clinic for the treatment of ALS. This review covers the VEGF family, its receptors and neurotrophic effects as well as VEGF therapy in animal models of ALS and advances towards clinical trials.

Expression of hypoxia inducible factor-1α and vascular endothelia growth factor in vocal polyps.

OBJECTIVES/HYPOTHESIS: Long-term vocal overuse or sudden vocal-fold microvascular disruption may lead to the formation of a hypoxic zone of injury and the subsequent release of a cascade of cytokines in the vocal fold. To test this hypothesis, we investigated the expression and the role of hypoxia inducible factor-1α (HIF-1α) and vascular endothelia growth factor (VEGF) in the formation of vocal polyps. METHODS: Expression patterns of HIF-1α and VEGF in surgical specimens of vocal polyps from 36 patients were analyzed using the immunohistochemistry and Western blot method. Normal vocal-fold mucosa from 26 patients who underwent total laryngectomy were collected and used as the control group. RESULTS: The expression of HIF-1α and VEGF were detected both in the vocal polyps group and the control group, while the expression of HIF-1α and VEGF levels were significantly higher when compared to normal vocal-fold mucosa (P < .05). CONCLUSION: The increased expression of HIF-1α in vocal polyps indicates that vocal fold overvibration induced hypoxia may play an important role in the pathogenetic mechanism of vocal polyps.

The protective effect of Clostridium butyricum on intestinal injure in newborn rats with necrotizing enterocolitis / 临床儿科杂志

Objective To explore the influence of Clostridium butyricum on the expression of vascular endothelial cell growth factor (VEGF) and its receptor 2 (VEGFR-2), proliferating cell nuclear antigen (PCNA), and tight junction protein claudin-2 in intestinal tissue in newborn rat with necrotizing enterocolitis (NEC). Methods Forty-eight-hour-old Sprague-Dewley (SD) rats were randomly divided into model group, control group, low-dose group, mid-dose group, and high-dose group, 12 rats each. Rats in each group were fed with milk substitute. The NEC model were created by hypoxia and cold stimulation for 3 consecutive days in model group, low-dose group, mid-dose group, and high-dose group. Meanwhile, low-dose group, mid-dose group, and high-dose group were intervened by being fed with Clostridium butyricum 0.2, 0.4, and 0.8 g/(kg·d), respectively. All rats in each group were sacriifced on day 4 and the intestines tissue was obtained. The pathological changes had been observed. The expression of VEGF, PCNA, and claudin-2 were detected by immunohistochemistry. The expression of VEGFR-2 was detected by RT-PCR. Results The intestines pathological scores was signiifcantly different among ifve groups (P?0 . 05 ). Conclusion The expression of VEGF, VEGF-2 , and claudin-2 were higher in rats with NEC, while the expression of PCNA was lower. Supplementation of Clostridium butyricum may protect newborn rats by its act on these factors.

Comparison on serum biomarkers for anovulatory and ovulatory dysfunctional uterine bleeding in Lizu females

OBJECTIVE@#To screen, identify, and compare the serum biomarkers between anovulatory dysfunctional uterine bleeding (ADUB) and ovulatory dysfunctional uterine bleeding (ODUB) in Lizu females.@*METHODS@#The subjects included 128 ADUB patients, 63 ODUB patients, and 93 controls. The serum and supernate of the subjects' mense were collected and stored at -80 °C until use. Differential proteins in the sera of three groups were screened using surface-enhanced laser desorption ionization time-of-flight mass spectrometry. The screened proteins were then identified by tricine-SDS-PAGE gel and spectrometry. Protein expression levels in the menses of ADUB, ODUB, and control subjects were determined using ELISA, RT-PCR, and Western blotting. SPSS 14.1 was used for statistical analysis and chart drawing (α = 0.05).@*RESULTS@#Three differential protein peaks with peak values of 11.80, 13.59, and 14.68 km/z were screened and identified as serum amyploid protein A (SAA), vascular endothelial growth factor, and vitamin K epoxide reductase, respectively. The SAA was highly expressed in the menses of ADUB and ODUB patients but poorly expressed in the controls. The vascular endothelial growth factor was highly expressed in the menses of ODUB and controls but poorly expressed in ADUB patients. Meanwhile, the vitamin K epoxide reductase was highly expressed in the menses of ADUB and control subjects but poorly expressed in ODUB patients.@*CONCLUSIONS@#The SAA is the common serum biomarker of ADUB and ODUB. ADUB may be related to angiogenesis impairment, whereas ODUB may be associated with blood coagulation disruption.

Effect of canonical transient receptor potential channels on hypoxia-induced VEGF expression in glioblastoma cells / 中华神经医学杂志

Objective To investigate the effect of canonical transient receptor potential (TRPC)channels on hypoxia-induced VEGF expression in U87 glioblastoma cells.Methods U87 cells cultured in vitro were divided into normoxia group,normoxia+25 μmol/L SKF96365 treatment group,normoxia+40 μmol/L SKF96365 treatment group,hypoxia group,hypoxia+25 μmol/L SKF96365 treatment group and hypoxia+40 μmol/L SKF96365 treatment group; 34 h after the cell culture,25 and 40 μmol/L SKF96365 were added into the treatment groups respectively,and hypoxia group and normoxia group were added the same amount of solvent; two h later,the medium in the later three groups were changed into the hypoxia ones.VEGF mRNA expression at each group was detected by real-time PCR after 10 h of hypoxia; Real-time PCR and Western blotting were used to observe the mRNA and protein expressions of TRPC1 in the hypoxia group and normoxia group after 16 h of hypoxia; the changes of TRPC location in U87 cells 1 h after hypoxia were observed by immunofluorescence staining.Results VEGF mRNA expression showed significant difference between each two groups (F=101.362,P=0.000); that in the hypoxia group was obviously increased as compared with that in the normoxia group (P＜0.05); as compared with that in the hypoxia group,VEGF mRNA expression in the hypoxia+25μmol/L SKF96365 treatment group was significantly decreased (P＜0.05),and that in the hypoxia+25μmol/L SKF96365 treatment group was obviously higher than that in the hypoxia+40 μmol/L SKF96365 treatment group (P＜0.05).TRPC1 mRNA and protein expressions in the hypoxia group showed no significant difference as compared with that in the normoxia group (t=0.493,P=0.648; t=0.490,P=0.650).Immunofluorescence staining showed that TRPC1 mainly located in the cell cytoplasm and membrane before hypoxia,while translocation of TRPC1 into plasma membrane was noted 1 h after hypoxia.Conclusion Translocation of TRPC1 into plasma membrane is involved in hypoxia-induced VEGF expression.

Expression of Nestin Protein and Its Relation with Vascular Endothelia Growth Factor in Brain of Neonatal Rats with Hypoxia-Ischemia Brain Damage / 实用儿科临床杂志

Objective To explore the relationship between the expression of nestin protein and the expression of vascular endothelia growth factor(VEGF)in hypoxia-ischemia brain damage(HIBD)of neonatal SD rats.Methods Fifty-six 7-day postnatal SD rats were randomly divided into 2 groups,sham operation(n=28)and hypoxic ischemic(HI)group(n=28).Animals were killed in 0,3,12,24 h,3,7,14 d after HIBD rat models established.The brain tissue was obtained to make paraffin section.Expressions of nestin and VEGF protein were examined with immunohistochemical staining and image quantitative analysis.Results The number of VEGF positive cell in the hippocampus and cortex increased reactively after HI and reached peak at the 12th hour and persistent to the 24th hour,which returned the normal level on d3,however,the number of nestin positive cell reached peak on d7 and returned the normal level on d14 after HI.Conclusions VEGF and nestin may contribute to neurolproctetive effects against HIBD.Studying the way to facilitate the coexpression of the nestin and VEGF may have important rolls to enhance the repairment and regeneration.

A study on biological activity of co-expression plasmid of human tissue plasminogen activator and vascular endothelial growth factor 165 / 中国普通外科杂志

Objective To observe the co-expression plasmid of tissue plasminogen activator (tPA) and vascular endothelia growth factor165 (VEGF165) in vascular endothelial cell (VEC) and to study the effect of the product on the proliferation of VEC and fibrinolysis activity. Methods pBudCE4.1/tPA-VEGF165 was transfected into VECs by using lipofection. The expression of tPA and VEGF165 at mRNA level was detected by RT-PCR and expression at protein level was detected by Western blot. The fibrinolysis activity of VEC culture solution of transfecting tPA and VEGF165 genes were detected by fibrin plate technique. The VEC and VSMC were cultured with VEC culture solution of transforming tPA and VEGF165 genes, the proliferation of VEC and VSMC were evaluated with 3?H-TdR incorporation and flow cytometry (FCM). Results The expression of tPA and VEGF165 in the transfected VECs was detected. The fibrinolysis activity of transfected VEC culture solution was also detected. tPA and VEGF165 products in VECs elevated proliferation of VEC, while there was no effect on the proliferation of VSMC. Conclusion The tPA and VEGF165 eukaryotic co-expression plasmid could express in transfected VECs, and the expression products have biology activity.

Study of expression of vascular endothelial growth factor in granulation tissue of burn wound and post-burn hypertrophic scar at excessive stages / 中国康复理论与实践

@#ObjectiveTo explore the role of vascular endothelial growth factor(VEGF) in the growth and development of hypertrophic scar.MethodsThe burn wound samples of various stages were selected from transition of wound granulation tissue to scar and in long-persisting post-burn hypertrophic scar, and the concentrations of VEGF protein were detected using enzyme-linked immunosorbent essay (ELISA) method. ResultsThe tissue homogenate concentration of VEGF protein increases gradually from the wound granulation tissue to hypertrophic scar before it achieves summit concentration during 4 to 6 month. The concentration of VEGF degreases gradually after the maturation of hypertrophic scar. The high concentration of VEGF is synonymous with the large amount of capillary of the immature scar.ConclusionsThe abnormal expression of VEGF is related to the growth and development of hypertrophic scar and induces excessive and uncontrollable angiogenesis.