Differences In Vascular Endothelial Growth Factor Expression Research Articles

Platelet Rich Fibrin as a Potential Treatment for Corneal Neovascularization due to Infective Keratitis: An Experimental Study

Abstract
 Introduction & Objectives : Platelet-rich fibrin (PRF) as a source of growth factor has been widely used in various therapeutic fields so that it could accelerate the wound healing process. This study investigated the effect of platelet-rich fibrin PRF in inhibiting neovascularization, inhibiting the expression of vascular endothelial growth factor (VEGF) and stimulating pigment epithelial-derived factor (PEDF) in the infective keratitis animal model.
 Methods : A total of 12 New Zealand white rabbits (Oryctalogus cuniculus) were subjected to keratitis induction by inoculation of Pseudomonas aeruginosa. The rabbits were divided into two groups, the first group as control was treated antibiotic-only, and the second as treatment group was treated with antibiotic and PRF graft. The corneal neovascularization area was clinically measured before and after treatment. Expression of VEGF and PEDF in the corneal epithelium, stroma, and endothelium were evaluated by immunohistochemical examination on day 10 after keratitis induction.
 Results : There were significant differences in VEGF expression between groups in the epithelial layer (p=0.008), but no significant difference in the stromal (p=0.934) and endothelial (p=0.935) layers. PEDF expression showed a significant difference between groups in the epithelial (p = 0.004) and stroma layers (p = 0.029) but no significant difference in the endothelial layer (p = 0.157). However, there was no significant difference in clinical neovascularization between the two groups (p=0.526).
 Conclusion : PRF could inhibit VEGF expression and stimulate PEDF expression in the cornea of Pseudomonas aeruginosa keratitis animal model that potentially reduces the corneal neovascularization in the longer observation.

The effects of prophylactic tadalafil use on VEGF expression in the rabbit model of steroid-induced femoral head avascular necrosis.

The purpose of this study was to examine the effect of prophylactic tadalafil use on a steroid-induced femoral head avascular necrosis model in terms of microscopic, imaging, and molecular biological changes. Twenty-four New Zealand rabbits were divided into 3 equal groups. Eight rabbits were designated as the control group and did not receive treatment. Rabbits in group 1 (G1) received 0.1 mg/kg Escherichia coli lipopolysaccharide (LPS) intravenously and 40 mg/ kg methylprednisolone sodium succinate (MP) was administered intramuscularly for 3 days consecutively. Rabbits in group 2 (G2) were given 5 mg/kg tadalafil orally for 10 consecutive days. Starting on the eighth day, 0.1 mg/kg LPS was given, and following this 40 mg/kg MP injections were administered for 3 days. All animals were sacrificed 3 weeks after the final MP injection. Magnetic resonance imaging was performed, and bilateral femora were harvested. Half of the femoral head was stored for Vascular Endothelial Growth Factor (VEGF) examination with Western blot analysis. The other half was examined microscopically for the presence of osteonecrosis. In G1, 15 out of 16 hips (93%) of the 8 rabbits had osteonecrosis compared to 8 out of 12 hips (67%) of 6 rabbits in G2 (P > .05). The VEGF expression in G2 was significantly higher than in the control group and G1 (P < .05 and P < .001, respectively). There was no significant difference in VEGF expression between the control group and G1 (P > .05). This study has shown us that femoral head osteonecrosis can be reliably induced with LPS and corticosteroid, as described in the literature. Prophylactic tadalafil use did not decrease the occurrence of osteonecrosis significantly. However, it significantly increased VEGF expression in the femoral head independent of the effects of steroids and LPS.

The effect of ropivacaine infiltration on the expression of Vascular Endothelial Growth Factor (VEGF) on skin defect: experimental study on Wistar rats

Link of Video Abstract: https://youtu.be/jNHUCHSk70Y Background: The main problems of postoperative patients are pain and wound. The healing phases consist of hemostasis, inflammation, proliferation, and remodeling. Postoperative pain causes surgical stress, which disrupts wound healing by delaying the transition from the inflammatory phase to the proliferative phase. Ropivacaine infiltration is expected to reduce surgical stress. One of the parameters that can be used to assess wound healing is Vascular Endothelial Growth Factor (VEGF) expression. This study aims to evaluate the infiltration effect of ropivacaine on the expression of VEGF in post-surgical skin defects in Wistar rats. Methods: This study was an experimental study using 32 Wistar rats (Rattus norvegicus), which were randomly divided into 2 groups. With a 2 cm full-thickness wound incision, the ropivacaine group was given 0.2% 1 ml of ropivacaine infiltration, while the control group was given NaCl infiltration. 0.9% 1 ml. Tissue samples were taken on day 5 to calculate VEGF expression through immunohistochemical examination. Data were analyzed using SPSS versin 23.0 for Windows. Results: Assessment of the weight of the Wistar rats did not show a significant difference (p = 0.788). Using the IRS (Immunoreactivity Score), there was a significant difference in VEGF expression (p = 0.033) between the two groups. There were significant differences in the degree of VEGF cells between the two groups (p = 0.001). Analysis of the relationship between treatment and degree of VEGF cells showed a significant relationship (p = 0.005). Conclusion: Infiltration of ropivacaine had effects on the expression of VEGF in postsurgical skin defects of Wistar rats

Effect of tumor-stromal fibroblasts on the biological behavior of salivary gland pleomorphic adenoma cells in vitro.

This study aims to investigate the effects of tumor-stromal fibroblasts (TSFs) on the proliferation, invasion, and migration of salivary gland pleomorphic adenoma (SPA) cells in vitro. Salivary gland pleomorphic adenoma cells (SPACs), TSFs, and peri-tumorous normal fibroblasts (NFs) were obtained by tissue primary culture and identified by immunocytochemical staining. The conditioned medium was obtained from TSF and NF in logarithmic phase. SPACs were cultured by conditioned medium and treated by TSF (group TSF-SPAC) and NF (group NF-SPAC). SPACs were used as the control group. The proliferation, invasion, and migration of the three groups of cells were detected by MTT, transwell, and scratch assays, respectively. The expression of vascular endothelial growth factor (VEGF) in the three groups was tested by enzyme linked immunosorbent assay (ELISA). Immunocytochemical staining showed positive vimentin expression in NF and TSF. Results also indicated the weak positive expression of α-smooth muscle actin (SMA) and fibroblast activation protein (FAP) in TSFs and the negative expression of α-SMA and FAP in NFs. MTT assay showed that cell proliferation in the TSF-SPAC group was significantly different from that in the NF-SPAC and SPAC groups (P<0.05). Cell proliferation was not different between the NF-SPAC and SPAC groups (P>0.05). Transwell and scratch assays showed no difference in cell invasion and migration among the groups (P>0.05). ELISA showed that no significant difference in VEGF expression among the three groups (P>0.05). TSFs may be involved in SPA biological behavior by promoting the proliferation of SPACs but has no effect on the invasion and migration of SPACs in vitro. Hence, TSF may be a new therapeutic target in SPA treatment.

The Effect of The Topical Application of Eurycoma Longifolia Jack (TA) Root Extract Hydrogel on Vascular Endothelial Growth Factor (VEGF) Expression During Wound Healing in vivo Excisional Wound Model

INTRODUCTION: Medicinal plants are known for their positive impacts on wound healing by stimulating angiogenesis in the skin. Eurycoma longifolia Jack locally known as Tongkat Ali (TA), is a medicinal plant characterised by anti-inflammatory and antioxidant effects. MATERIALS AND METHODS: Excisional wound (15mm × 15mm) in diameter and (2mm) depth was created at the back of 20 male Sprague Dawley rats by incising the marked skin with sterilized surgical scalpel blade then excised the skin by surgical scissors and toothed forceps. The wounded rats were divided into 4 groups, each group contained 5 rats (n=5). Experimental groups were formed as follows: untreated (-ve) control, Hydrocyn® aqua gel (+ve) control, vehicle hydrogel, and TA hydrogel. All the treatments were applied twice daily for 5 days starting on the first day (wounding day). On Day 5 post-wounding, the granulation tissue was harvested from all groups and evaluated by immunohistochemistry assay for Vascular endothelial growth factor (VEGF) expression. RESULTS: VEGF expression in granulation tissue of rat’s skin treated with TA hydrogel and vehicle hydrogel increased significantly compared to that in the untreated (-ve) control group with p-values of 0.040 and 0.029, respectively. Although there was no significant difference in VEGF expression in granulation tissue of rat’s skin treated with the TA group, Hydrocyn® aqua gel (+ve) control, and vehicle hydrogel groups, our study group showed higher expression of VEGF. CONCLUSION: Our study showed that the topical application of TA hydrogel increased the VEGF expression in granulation tissue of rat’s skin, which is an essential growth factor for wound healing. Thus, there is great potential for TA hydrogel to be an effective wound-healing agent for managing cutaneous wounds.

Fibrin glue: an anti-fibrotic agent for contracted socket

Background: The contracted socket is caused by fibrosis leading to the shortening of the orbital tissue. Vascular endothelial growth factor (VEGF) is a profibrotic factor in postoperative wound healing. The novel option to prevent fibrosis is the use of fibrin glue. This study aims to evaluate the role of fibrin glue in reducing the VEGF expression in cells that cause post-socket surgery scar. Methods: An in vivo experimental study using an animal model was conducted. We used twenty eyes of white rabbit species as the study sample. The evisceration was carried out and distributed to the animals in four experimental groups consisting of the control group, the MMC group (0.1 ml of mitomycin-C (MMC) 0.4 mg/ml), the triamcinolone acetonide (TCA) group (0.1 ml of TCA 40 mg/ml), and the fibrin glue group (0,1 ml of fibrin glue). The treatment was performed through injection of the subconjunctival. After 14 days, rabbits were euthanized, and conjunctiva samples were cut. Immunoreactivity scores (IRS) were used to analyze the expression of VEGF in epithelia conjunctiva with a significant level of p &lt;0.05. Data were analyzed using SPSS version 26 for Windows. Results: All groups showed a statistically significant difference in VEGF expression with a p-value 0,000 (p&lt;0,05). The VEGF level was significantly lower in the three treatment groups compared with the control group. The VEGF expression was highest for the control group, followed by fibrin glue, TCA and MMC group. We found significant differences between the control group and the three treatment groups; the MMC group (p=0.001), the TCA group (p=0.002), and the fibrin glue group (p=0.005). While differences in VEGF expression between the three treatment groups could not be proven statistically. Conclusion: Fibrin glue is proven to reduce VEGF expression as a marker of fibrosis; thus, fibrin glue can be an alternative anti-fibrotic agent.

Mast cell degranulation and vascular endothelial growth factor expression in mouse skin following ionizing irradiation

ABSTRACTThe present study aimed to identify the mechanisms underlying the increase in vascular permeability in mouse skin following irradiation. The left ears of C3H mice were subjected to 2 and 15 Gy of radiation in a single exposure. At 24 h after irradiation, the ears were excised and tissue sections were stained with toluidine blue to assess mast cell degranulation. Vascular endothelial growth factor (VEGF) expression was assessed via immunohistochemistry and western blotting. Approximately 5% (3%–14%) (mean [95% CI]) of mast cells in the skin of control mice were degranulated; moreover, at 24 h after 2 Gy irradiation, this value increased to approximately 20% (17%–28%). Mast cell degranulation by 15 Gy irradiation (32% [24%–40%]) was greater than that by 2 Gy irradiation. Significant differences were observed in mast cell degranulation among the control, 2 Gy and 15 Gy groups (p = 0.012). Furthermore, VEGF-positive reactions were observed in the cytoplasm of scattered fibroblasts in the dermis. In immunohistochemistry tests, VEGF expression at 24 h after irradiation increased slightly in the 2 Gy group compared to that in the control group, whereas no difference in VEGF expression was observed in the 15 Gy group compared to that in the control group. Expression of VEGF in western blots was consistent with that in immunohistochemistry.In conclusion, mast cell degranulation was increased in mouse skin at 24 h after irradiation in a dose-dependent manner. In contrast, VEGF expression was slightly increased following only low-dose (2 Gy) irradiation.

ML-01 Pathological characteristics of primary central nervous system lymphoma with atypical radiological finding

Background: If the brain tumor is suspected to be a primary central nervous system lymphoma (PCNSL) on radiological findings, it is general to perform biopsy to obtain the pathological diagnosis. Glioblastomas (GBs) must be distinguished from PCNSLs. In addition to commonly used contrast-enhanced T1-weighted imaging, diffusion-weighted image (DWI), and apparent diffusion coefficient (ADC) value, the following characteristics of PCNSLs were reported to be essential for this purpose: 1) no increase in blood flow on perfusion images obtained by the arterial spin labeling (ASL) method; 2) less microbleeding on T2*-weighted images (T2*). However, we experienced some exceptional cases. Purpose: To clarify the histopathological features of PCNSLs those had atypical radiological findings. Method: 62 consecutive PCNSL cases (40 males, 22 females, mean age 65.4 years, range 35–84) treated in our department from April 2013 to March 2020 were retrospectively analyzed. We compared the following features between 47 biopsy cases showing typical image findings as PCNSLs (Group A) and 15 surgically resected cases with atypical findings (Group B), 1) number of blood vessels per hyper 10 fields, 2) occupying area of blood vessels per unit area, 3) immunoreactivity of vascular endothelial growth factor (VEGF), and 4) germinal center B-cell (GCB) subtype. Results: In Group A, the number of blood vessels in the tumor was 39.3 on average, and the area occupied by blood vessels was 3.8%. In Group B, the former was 133.2, and the latter was 9.9%. There was no significant difference in VEGF expression and GCB subtype. Conclusion: In PCNSLs showing with high blood flow and microbleeds, the blood vessels were rich and partial bleeding was confirmed histologically. We should analyze much more cases to set the threshold both of the ADC value and the absolute value of blood flow calculated by the ASL method to distinguish between PCNSLs and GBs.

The Expression of VEGF and CD31 in Endometrial Lesions and Its Associations with Blood Flow Parameters of Transvaginal 3D Power Doppler Ultrasonography: A Preliminary Study.

AimTo investigate the association of the blood flow parameters measured by transvaginal three-dimensional power Doppler ultrasound and the angiogenesis of endometrial cancer.Material and MethodsThe expressions of vascular endothelial growth factor (VEGF) and CD31 in benign and malignant endometrial lesions, and in malignant lesions with different clinical and pathological features were analyzed. The correlations of the blood flow parameters (vascularization index [VI], blood flow index [FI], and vascularization-blood flow index [VFI]) of transvaginal 3D power Doppler ultrasound, and VEGF expressions, microvessel density (MVD) were also evaluated.ResultsThe VEGF-positive rates and the MVD values in benign and malignant endometrial lesions were significantly different (both P<0.001). The differences of VEGF-positive rates (P < 0.001) and MVD values (P = 0.021) between type I and type II lesions of endometrial cancer were statistically significant. There was no significant difference in the VEGF-positive rate and MVD value between stage IA and IB (P=0.443, P=0.311). The difference of VEGF expression and MVD in stage IA, stage IB and stage II and above was statistically significant (P=0.003, P=0.017). The VEGF-positive rate and MVD value were not significantly different in IAG1 and IAG2 lesions of endometrioid adenocarcinoma (P=0.709, P=0.792). There was no significant correlation between VI, FI, VFI and VEGF expression and MVD in endometrial cancer.ConclusionThe VEGF-positive rates and MVD values were relatively high in malignant endometrial lesions, type II and stage II and above lesions of type I endometrial cancer, indicating that the angiogenesis of endometrial cancer tissues might play a crucial role in the tumor classification, and pelvic metastasis.

VEGF promotes migration and invasion by regulating EMT and MMPs in nasopharyngeal carcinoma.

Background: Vascular endothelial growth factor (VEGF) is an important pro-angiogenic factor. Accumulating data have indicated that VEGF is involved in tumour metastasis. However, the mechanism through which VEGF regulates nasopharyngeal carcinoma (NPC) metastasis is largely unknown. This study aimed to examine the biological function of VEGF in NPC metastasis and its underlying mechanism.Methods: We used western blotting and qPCR to examine the difference in VEGF expression between NPC cells and the immortalized nasopharyngeal epithelial cell line NP69. Wound healing assays, transwell assays and animal experiments were used to further verify the role of VEGF in the invasion and migration of NPC cells. The protein levels of the epithelial-mesenchymal transition (EMT) and matrix metalloproteinase (MMP) family were analysed by immunofluorescence (IF) and western blotting. Enzyme-linked immunosorbent assay (ELISA) and transwell assays were used to determine whether VEGF enhanced the invasion and migration of NPC cells in an autocrine manner. Western blotting was used to examine how autocrine VEGF-VEGFR2 signalling regulated EMT and MMPs.Results: We observed higher levels of VEGF in NPC cells than that in NP69 cells and identified an association between high VEGF levels and tumour invasion and migration. Mechanistically, the VEGF-mediated increase in EMT markers, MMP2 and MMP9 promoted NPC cell invasion and migration. Additionally, NPC cells secreted VEGF to promote cell invasion, migration and angiogenesis. Autocrine VEGF-VEGFR2 signalling increased ERK1/2 phosphorylation, promoted EMT process and MMPs at the indicated times.Conclusion: This study revealed that VEGF plays a role in controlling NPC cell metastasis by regulating EMT markers and MMPs in an autocrine manner.

Role of angiogenesis in oral submucous fibrosis using vascular endothelial growth factor and CD34: An immunohistochemical study.

Oral submucous fibrosis (OSF) is an insidious, chronic, disabling disease, in which there is lack of perfusion due to reduced level of the vasculature and this is said to be responsible for the epithelial atrophy seen in OSF. The degree of vasculature of the affected mucosa and its effects on the epithelial thickness remains controversial till date. This study attempts to analyze the role of angiogenesis in OSF and its progression using vascular endothelial growth factor (VEGF) and CD34 markers. The study samples for the present study comprised of 10 cases each of early OSF, moderately advanced, advanced OSF, and 10 cases of normal oral mucosa were used as controls. All the cases were subjected to immunohistochemical staining with VEGF and CD34 markers. Among the different grades of OSF, we did not find any noticeable difference in VEGF expression although we found a upregulation in microvessel density (CD34) in early and moderately advanced OSF followed by a downregulation in advanced OSF. As the disease progresses, there is an increased production of the extracellular matrix component (collagen I and II and fibronectin) and results in fibrosis. Subsequently, it leads to the reduction in the level of corium vascularity and results in hypoxia which ultimately causes reduction and constriction of the vascular channels. This sequence of events alerts us to the relevance of early disease diagnosis and management in an irreversible pathology such as OSF.

Histologic Evaluation of Leucocyte- and Platelet-Rich Fibrin in the Inflammatory Process and Repair of Noncritical Bone Defects in the Calvaria of Rats.

This study aimed to evaluate the effects of leucocyte- and platelet-rich fibrin (L-PRF) on the inflammatory process, tissue repair, and expression of vascular endothelial growth factor (VEGF) on bone defects in the calvaria of rats. L-PRF was obtained from three animals submitted to cardiac puncture to prepare the membranes. Two noncritical defects with a diameter of 2 mm were created in the calvaria of 15 Wistar rats. The defects on the right side were filled with a blood clot (CTRL) and the left side with L-PRF. After 5, 15, and 30 days, the animals were euthanized and the specimens processed for histologic, histomorphometric, and immunohistochemical analyses. In order to measure the intensity of the inflammatory infiltrate and VEGF expression, scores were assigned from 0 to 3, with 0 being no expression, 1 discrete (up to 25%), 2 moderate (between 25% and 50%), and 3 intense (> 50%) expression. The area of bone neoformation at the edges of the defects was also quantified. A less intense inflammatory infiltrate was observed in the defects filled with L-PRF compared with CTRL at all times analyzed (P < .05). At 5 days, no bone neoformation was observed in any of the groups evaluated. After 15 and 30 days, greater bone neoformation was observed in the group treated with L-PRF compared with the CTRL group (P < .05). At 15 days, 3,871.8 (1,070.15) μm2 were recorded for the CTRL and 49,978.5 (14,360.7) μm2 in the L-PRF. At 30 days, 62,284.5 (3,579.5) μm2 were observed in the CTRL and 154,076.6 (31,464.9) μm2 in the L-PRF. At all evaluated times, a lower inflammatory infiltrate was observed in the group treated with L-PRF compared with the CTRL. VEGF expression was observed in the initial phase and throughout the tissue repair process in both groups. At 5 days, there was no difference in VEGF expression between the groups. VEGF was present at the initial phase and throughout the tissue repair process in both groups. In the L-PRF group, a decrease in VEGF expression was observed at 15 and 30 days compared with the CTRL group. L-PRF had a positive effect on the regenerative process of bony defects, with a reduced inflammatory response and greater bone neoformation.

EFFECTS OF COWPEA (VIGNA UNGUICULATA) EXTRACT IN VEGF EXPRESSION OF CORNEAL INFLAMMATION RAT MODEL (RATTUS NOVERGICUS STRAIN WISTAR)

Purpose : to evaluate the effects of Methanolic extract of Cowpea (Vigna unguiculata) administration on vascular endothelial growthfactor (VEGF) expression in rats with corneal-alkali induced inflammation.Methods : This was an interventional experimental study. Sixty three rats were randomly selected, 3 rats as control, 60 rats were treatedwith 1 M NaOH (as inflammation model) and administered with 25 ìM,50 ìM and 100 ìM of genistein in Cowpea (Vignaungucuilata) extract four times daily. Alkali burn by 1 M NaOH infiltration using filter paper applicated on the center cornea of righteye for 60 seconds. Aquabidest as positive control and Cowpea (Vigna ungucuilata) extract treatment perfused immediately after alkali burn.VEGF observed at 6, 24, 48, 96 and 168 hours by immunohistochemical method.Results : Each dose of Cowpea (Vigna unguiculata) extract administration significantly decreased the VEGF expression ( p = 0,00). Differences of VEGF expression based on administration time were not significant (p=0,033). Interaction between time of administrationand dose of administration had influenced the VEGF expression (R Square(r2) =24,7 %). The Linear regression between VEGF expression and dose resulted in estimated effective dose (ED) with Y = 16.486–0.079 X( Y = V E G F e x p r e s s i o n , X =d o s e o f a d m i n i s t r a t i o n ) . Considering VEGF expression in normal cornea was 12, wefound the effective dose of methanolic Vigna unguiculata extract was 61.94 ìM.Conclusion :Methanolic extract of Cowpea(Vigna unguiculata) decreased the VEGF expression on alkali burn corneal inflammationin rats but no differences in VEGF expression based on time administration. Dose of 50 ìM genistein in Methanolic extract ofCowpea (Vigna unguiculata) was close to effective dose 61.94 ìM.Keywords : VEGF expression, alkali burn, Cowpea (Vigna unguiculata) extract, cornea.

Vascular endothelial growth factor expression in placenta of hypertensive disorder in pregnancy.

Hypertensive disorder in pregnancy (HDP) represents the most common medical complication in pregnancy. It is the leading cause of maternal and perinatal mortality and morbidity. Vascular endothelial growth factor (VEGF) stimulates vascular endothelial cell growth, survival, and proliferation, and they are known to be expressed in human placenta. The aim of this study was to determine the VEGF expression in the placenta of hypertensive and normotensive patients. This is a retrospective, cross-sectional study from January 1, 2015 to December 31, 2015. A total of 30 placentae comprised of 15 hypertensive and 15 normotensive cases were assessed. VEGF expression in placenta was assessed by immunohistochemistry, and the number of syncytial knots was counted. Our study showed an increased syncytial knot formation in the placenta of hypertensive mothers. VEGF expression was seen in syncytiotrophoblasts of 14 of the hypertensive cases (14/15, 93.3%), while only two of the normotensive cases were positive (2/15, 13.3%). There were no statistically significant differences in VEGF expression in other placenta cells, that is, cytotrophoblasts (P = 1.0), decidual cells (0.1394), maternal endothelial cells (0.5977), and fetal endothelial cells (P = 1.0). This study showed an increased number of syncytial knots is a consistent histological finding in the placenta of patient with HDP. VEGF expression was significantly increased in syncytiotrophoblasts in placenta of hypertensive group, and it could be used as a biomarker for hypertension.

Biomarkers of angiogenesis as prognostic factors in myelodysplastic syndrome patients treated with hypomethylating agents

Angiogenesis occurs in response to tissue ischemia and wound healing, and contributes to the pathogenesis of a variety of diseases, such as benign and malignant neoplasia. Several studies have measured bone marrow microvessel density (MVD) in MDS patients and acute myeloid leukemia (AML) patients transformed from MDS, and MVD was higher in MDS patients than controls, but was lower than in AML patients. Vascular endothelial growth factor (VEGF) is expressed in bone marrow blast cells, and an autocrine VEGF signaling mechanism has been established in MDS. Increased bone marrow angiogenesis and VEGF concentrations are adverse prognostic features in all of these patients. In this study, 69 patients were treated in two groups: hypomethylating agents or supportive care with oxymetholone±pyridoxine. We evaluated the MVD and VEGF expression of paraffin-embedded bone marrow samples from patients. We also investigated the relationship between angiogenesis-related biomarkers including MVD, VEGF expression, and clinical factors.The patient median age was 65 years, and the median follow-up duration was 28 months. MVD assessment among subtypes of WHO MDS classification showed that the MVD of RCUD was significantly lower than in other types (p=0.02). However, there was no significant difference in VEGF expression according to the subtype of MDS. Although MVD and VEGF expression did not differ between risk groups based on the IPSS, the low risk group tended to have lower expression of angiogenesis-related biomarkers. MDS patients receiving hypomethylating agents had significantly lower MVD expression in responders than in non-responders (6.13±3.38 vs. 9.89±2.10, respectively, p=0.039). In a consecutive evaluation at the time of diagnosis and 3 months after the initial treatment, the group with a decrease or no change of MVD had a higher response rate compared to that in the group with an increase of MVD (92.9% vs. 58.8%, respectively, p=0.045). Adverse prognostic factors included older age, MDS type other than RCUD, a higher IPSS risk group, and abnormal cytogenetics. Although angiogenesis-related markers did not demonstrate any significant prognostic association with survival, MVD (≥10n/mm2) and a strong expression of VEGF seemed to be associated with lower survival rate.These data suggested that the MVD value might be helpful in predicting responsiveness to treatment, especially in MDS patients treated with hypomethylating agents. Although angiogenesis-related markers including VEGF did not demonstrate a significant association with survival outcomes, we observed that high MVD and strong VEGF expression seemed to be associated with lower survival rate. Therefore, biologic markers related to angiogenesis might have a potential as prognostic factors for MDS patients.

Relationship between vascular endothelial growth factor and microvessel density in placenta and placenta increta

Objective To investigate the relationship between the expression level of vascular endothelial growth factor (VEGF) and microvessel density (MVD) in placenta tissue and placenta increta. Methods Thirty singleton pregnant women who received antenatal care and underwent cesarean section in Fujian Maternity and Child Health Hospital between November 2013 and August 2014, were enrolled in this study. They were divided into placenta previa group, placenta increta group and control group, with ten patients in each group. Placenta tissue was collected from each patient. Expressions of VEGF and its mRNA were detected by immunohistochemistry, Western blot, and real-time polymerase chain reaction. MVD in placenta tissue was measured by immunohistochemistry. Rank sum test, t test, Kruskal Wallis test, one-way ANOVA and Spearman correlation test were used for statistical analysis. Results (1) Gestational age at admission and delivery in placenta previa and placenta increta groups was lower than in control group (all P<0.05). Compared with control group, the placenta previa and placenta increta groups had more blood loss, and longer operating duration and hospital stay (all P<0.05). (2) The expression levels of VEGF and its mRNA in placenta increta and placenta previa groups were higher than in control group (VEGF: 0.691±0.032, 0.695±0.027 and 0.518±0.025, respectively, F=373.401; VEGF mRNA: 1.667±0.661, 1.832±0.678 and 0.767±0.269, respectively, F=27.399; both P<0.05). But there was no significant difference between placenta previa and increta groups. (3) There was no significant difference of VEGF expression in increta location, border site and normal site in placenta increta group, but its mRNA was decreasing (2.519±0.116, 1.482±0.232 and 1.000±0.000, respectively, F=240.827, P<0.05). (4) Expression level of VEGF at the attachment of umbilical cord, upper and lower margin of placenta in placenta previa group was higher than in control group (0.702±0.026 vs 0.528±0.020, t=12.302; 0.698±0.026 vs 0.519±0.035, t=12.715; and 0.685±0.029 vs 0.509±0.010, respectively, t=17.891; all P<0.05). Expression of VEGF mRNA in placenta previa group was higher than in control group (2.080±0.539 vs 1.024±0.272, t=8.093; 1.587±0.757 vs 0.546±0.083, t=2.401; 1.828±0.704 vs 0.731±0.157, t=4.259; all P<0.05). (5) MVD in placenta increta group and placenta previa group was higher than in control group (171.2±14.7, 155.7±14.6 vs 147.8±12.3, respectively, F=7.277, P<0.05). (6) Expression level of VEGF in placenta increta group and control group was positively associated with MVD (r=0.825, P<0.05). Conclusions There may be some common mechanisms in the occurrence of placenta previa and placenta increta. Overexpression of VEGF in placenta and abnormal formation of villous vessels may be important factors in the pathogenesis of placenta increta. Key words: Placenta accreta; Placenta previa; Vascular endothelial growth factor A; Placenta; Microvessels

Clinicopathologic Characteristics of Synchronous Primary Thyroid Cancer Detected by Initial Staging 18F-FDG PET-CT Examination in Patients with Underlying Malignancy

The objective of this study was to define clinicopathologic characteristics in concurrent primary thyroid cancer detected by initial (18)fluorine-fluorodeoxyglucose positron emission tomography-computed tomography ((18)F-FDG PET-CT) scanning in patients with underlying malignancy. Among 155 patients with known underlying malignancy and with focal FDG uptake in the thyroid, 25 patients (22 females; mean age ± SD 54.4 ± 11.2 years; age range 27-70 years) who were confirmed as having papillary thyroid cancer (PTC; synchronous thyroid cancer) by cytological examination were included. Another 25 patients (24 females; mean age ± SD, 48.8 ± 12.7 years) with focal uptake in preoperative (18)F-FDG PET-CT due to PTC and no history of other malignancy (primary thyroid cancer) were also included. Immunohistochemical studies were performed for glucose transporter-1 (GLUT-1) and vascular endothelial growth factor (VEGF). GLUT-1 expression was significantly lower in synchronous thyroid cancer (7 of 25 patients, 28%) compared with primary thyroid carcinoma (15 of 25 patients, 60%; p = 0.045). However, age and tumor size of synchronous thyroid cancer were not significantly different from the patients with primary thyroid carcinomas. There was no significant difference in VEGF expression, maximal standardized uptake values, extrathyroidal extension, lymph node metastasis, advanced stage, and multifocality between both thyroid cancer groups. Clinicopathologic characteristics of synchronous thyroid cancer in patients with underlying malignancy were not different from those of patients with primary thyroid cancers except for GLUT-1 expression.

Clinicopathologic features and expression of epidermal growth factor receptor and vascular endothelial growth factor in adrenocortical tumors

To study the clinicopathologic features and expression of epidermal growth factor receptor (EGFR) and vascular endothelial growth factor (VEGF) in adrenocortical tumors. Forty-two cases of adrenocortical tumors operated at the Beijing Union Medical College Hospital during the period from July, 2001 to July, 2010 were retrospectively reviewed. Immunohistochemical study for EGFR and VEGF was carried out. The clinical information and follow-up data were analyzed. The cases included 21 adrenocortical carcinomas (ACC) and 21 adrenocortical adenomas (ACA). Nine patients suffered from primary aldosterone syndrome, including 8 cases with ACA and 1 case with ACC. The average tumor size, tumor weight, and duration between disease onset and diagnosis in the 21 cases of ACC were 11.7 cm, 542 g and 8.5 months, respectively. This was in contrast to 3 cm, 9.8 g and 45.6 months, respectively in cases of ACA. Histologically, the WEISS score in all the 21 cases of ACA was ≤ 2 (average = 0.9). None of the ACC cases had score less than 4 (average = 6.6). The presence of sinus invasion correlated with tumor metastasis (P < 0.01). Immunohistochemical study showed that EGFR was expressed in 61.9% of ACC patients (13/21), whereas EGFR staining was mostly negative in ACA (except for weak staining in 5 cases and moderate staining in 1 case). The difference of EGFR expression between ACC and ACA was statistically significant (P = 0.030). On the other hand, the positive rate of VEGF in ACC was 71.4% (15/21), including 28.6% (6/21) with strong expression and 28.6% (6/21) with moderate expression. In contrast, the expression rate of VEGF in ACA was 30.0% (7/21), including 14.3% (3/21) with moderate expression. The difference of VEGF expression between ACC and ACA was statistically significant (P = 0.013). There was correlation between VEGF expression and venous invasion (P = 0.028). The average duration of survival in patients with ACC was shorter than that in ACA. The tumor weight in ACC also correlated with prognosis. Tumor size, weight and presence of endocrine symptoms may help in the differential diagnosis between ACC and ACA. A WEISS score of ≥ 3 highly suggests ACC. The presence of sinus invasion is associated with metastasis. EGFR or VEGF expression may also be important in differentiating ACC from ACA.

Effects of estrogen replacement therapy on apoptosis and vascular endothelial growth factor expression in ocular surface epithelial cells: An experimental study.

To investigate the effects of estrogen replacement therapy (ERT) on apoptosis and vascular endothelial growth factor (VEGF) expression in ocular surface in an experimental rat model. Forty female, Wistar rats were randomized in 4 groups in the study. Subcutaneous ERT (17β-estradiol, 10µg/kg/day) was administered to the first group without ovariectomy and to the second group with ovariectomy for three months. Third group had only ovariectomy and fourth group had sham operation. All rats were sacrificed in estrous cycles determined by vaginal smear test and their right eyes were enucleated at the end of the third month. Enucleated eyes were analyzed by immunohistochemical method for expressions of caspase-3, bcl-2, VEGF and TUNEL assay. Caspase-3 expression of conjunctival epithelium was significantly higher in group 3 than group 1 (P=0.005), and group 2 (P=0.007). TUNEL score of conjunctival epithelium was significantly higher in group 3 than group1 (P=0.006). TUNEL score of corneal epithelium was significantly higher in group 3 than group 2 (P=0.012), and group 4 (P=0.002). There was no significant difference between groups in that bcl-2 and VEGF expressions. We determined increased apoptosis in ocular surface epithelial cells in ovariectomized rats. ERT and endogen estrogen decreased the apoptosis, and did not result in difference in VEGF expression between the groups. Estrogen may be beneficial for the treatment of apoptosis-mediated ocular surface disorders such as dry eye. Further studies are needed on this subject for a better understanding of the role of estrogen and to provide a new insight for treatment and prevention of apoptosis-mediated ocular surface disorders.

Angiogenesis and Survival in Patients with Myelodysplastic Syndrome

Angiogenesis has been implicated in the pathogenesis and prognosis of myelodysplastic syndrome (MDS). In this study, we investigated the relationship between microvessel density (MVD), vascular endothelial growth factor (VEGF) expression, common morphological and clinical factors, and survival in patients with MDS. We examined the MVD of paraffin-embedded bone marrow sections from 70 MDS patients and 31 controls. VEGF expression was determined in 50 patients and 20 controls. The median MVD in MDS patients was significantly higher than that in controls (p = 0.025), whereas there was no difference in VEGF expression between MDS patients and controls. In univariate analysis, increased MVD was associated with a shorter survival time (p = 0.023). However, in multivariate analysis, MVD was not an independent predictor of survival. The VEGF expression did not influence survival in univariate analysis. Survival was independently influenced by platelet count (p = 0.0073), cytogenetic risk category (p = 0.022), and transfusion dependence (p = 0.0073). Neither MVD nor VEGF expression were predictors for progression to acute myeloid leukemia in univariate analysis. Progression to acute myeloid leukemia was independently influenced only by the cytogenetic risk category (p = 0.022). This study confirmed increased MVD in MDS. It does not support an independent prognostic role of angiogenesis in MDS.