Expression of vascular endothelial growth factor and vascular endothelial growth factor receptors 1 and 2 in invasive breast carcinoma: prognostic significance and relationship with markers for aggressiveness

Dhakal H P, Naume B, Synnestvedt M, Borgen E, Kaaresen R, Schlichting E, Wiedswang G, Bassarova A, Holm R, Giercksky K‐E & Nesland J M
(2012) Histopathology  61, 350–364 Expression of vascular endothelial growth factor and vascular endothelial growth factor receptors 1 and 2 in invasive breast carcinoma: prognostic significance and relationship with markers for aggressiveness Aims: Vascular endothelial growth factor (VEGF), VEGF receptor 1 (VEGFR‐1) and VEGF receptor 2 (VEGFR‐2) play a role in breast cancer growth and angiogenesis. We examined the expression and relationship with clinical outcome and other prognostic factors. Methods and results: Tumour sections from 468 breast cancer patients were immunostained for VEGF, VEGFR‐1, and VEGFR‐2, and their relationships with tumour vascularity, disseminated tumour cells (DTCs) in bone marrow and other clinicopathological parameters were evaluated. VEGF, VEGFR‐1 and VEGFR‐2 immunoreactivities were observed in invasive breast carcinoma cells. VEGF expression was significantly associated with VEGFR‐1 and VEGFR‐2 expression (P < 0.001). High‐level cytoplasmic expression of VEGFR‐1 was associated with significantly reduced distant disease‐free survival (DDFS) (P = 0.017, log‐rank) and breast cancer‐specific survival (BCSS) (P = 0.005, log‐rank) for all patients, and for node‐negative patients without systemic treatment (DDFS, P = 0.03, log‐rank; BCSS, P = 0.009, log‐rank). VEGFR‐1 expression was significantly associated with histopathological markers of aggressiveness (P < 0.05). Significantly reduced survival was observed in DTC‐positive patients as compared with DTC‐negative patients in the combined moderate/high VEGFR‐1 group (P < 0.001 for DDFS and BCSS), and the same was true for DDFS in the moderate VEGFR‐2 group (P = 0.006). Conclusions: High‐level expression of VEGFR‐1 indicates reduced survival. Higher‐level expression of VEGFR‐1 or VEGFR‐2 in primary breast carcinomas combined with the presence of DTC selects a prognostically unfavourable patient group.     

Expression and Cellular Distribution of Vascular Endothelial Growth Factor‐C System in Cortical Tubers of the Tuberous Sclerosis Complex

Cortical tubers are malformations of cortical development in patients with tuberous sclerosis complex (TSC), and highly associated with pediatric intractable epilepsy. Recent evidence has shown that signaling mediated through vascular endothelial growth factor‐C (VEGF‐C) and its receptors, VEGFR‐2 and VEGFR‐3, has direct effects on both neurons and glial cells. To understand the potential role of VEGF‐C system in the pathogenesis of cortical tubers, we investigated the expression patterns of VEGF‐C signaling in cortical tubers compared with age‐matched normal control cortex (CTX). We found that VEGF‐C, VEGFR‐2 and VEGFR‐3 were clearly upregulated in tubers at both the mRNA and protein levels, compared with CTX. The in situ hybridization and immunostaining results demonstrated that VEGF‐C, VEGFR‐2 and VEGFR‐3 were highly expressed in dysplastic neurons (DNs), giant cells (GCs) and reactive astrocytes within tubers. Most DNs/GCs expressing VEGF‐C and its receptors co‐labeled with neuronal rather than astrocytic markers, suggesting a neuronal lineage. In addition, protein levels of Akt‐1, p‐Bad and ERK1/2, the important downstream factors of the VEGF‐C pathway, were significantly increased in cortical tubers, indicating involvement of VEGF‐C–dependent prosurvival signaling in cortical tubers. Taken together, our results suggest a putative role for the VEGF‐C signaling pathway in the pathogenesis of cortical tubers.     

Expression of Matrix Metalloproteinase‐9 mRNA and Vascular Endothelial Growth Factor Protein in Gastric Carcinoma and Its Relationship to Its Pathological Features and Prognosis

To investigate matrix metalloproteinase‐9 (MMP‐9) mRNA and vascular endothelial growth factor (VEGF) protein expression in gastric carcinoma and its correlation with microvascular density, growth‐pattern, invasion, metastasis, and prognosis. In situ hybridization of MMP‐9 mRNA and immunohistochemistry of VEGF and CD34 proteins were performed on surgical specimens of gastric cancers from 118 patients compared with 20 nonmalignant gastric mucosae. Their relationships to pathological parameters and survival times were determined by statistical analysis. The positive rate of MMP‐9 in noncancerous gastric mucosae was significantly lower than that of gastric cancer tissue (60.17%, P < 0.01). In patients with cancers of the infiltrating type, at stage T3‐T4, with vessel invasion, lymphatic metastasis, hepatic, or peritoneal metastasis, the positive expression rates of MMP‐9 mRNA, VEGF protein, and CD34 were significantly higher than those for patients with tumors of the expanding type (P < 0.01), at stage T1–T2 (P < 0.01), with nonvessel invasion (P < 0.05), without lymphatic metastasis (P < 0.05), and without hepatic (P < 0.001) or peritoneal metastasis (P < 0.001), respectively. Expression of MMP‐9 mRNA was positively related to that of VEGF protein (P < 0.001) and microvascular density (P < 0.001). Patients with higher MMP‐9 mRNA and VEGF expression demonstrated vivid tumor angiogenesis and poor 5‐year survival rate. MMP‐9 and VEGF expression is associated with enhanced tumor angiogenesis and may play crucial roles in the invasion and metastasis of gastric carcinoma. Therefore, MMP‐9 and VEGF may represent prognostic biomarkers and promising targets for therapeutic intervention. Anat Rec, 2010. © 2010 Wiley‐Liss, Inc.     

Altered ratios of pro‐ and anti‐angiogenic VEGF‐A variants and pericyte expression of DLL4 disrupt vascular maturation in infantile haemangioma

Infantile haemangioma (IH), the most common neoplasm in infants, is a slowly resolving vascular tumour. Vascular endothelial growth factor A (VEGF‐A), which consists of both the pro‐ and anti‐angiogenic variants, contributes to the pathogenesis of IH. However, the roles of different VEGF‐A variants in IH progression and its spontaneous involution is unknown. Using patient‐derived cells and surgical specimens, we showed that the relative level of VEGF‐A₁₆₅b was increased in the involuting phase of IH and the relative change in VEGF‐A isoforms may be dependent on endothelial differentiation of IH stem cells. VEGFR signalling regulated IH cell functions and VEGF‐A₁₆₅b inhibited cell proliferation and the angiogenic potential of IH endothelial cells in vitro and in vivo. The inhibition of angiogenesis by VEGF‐A₁₆₅b was associated with the extent of VEGF receptor 2 (VEGFR2) activation and degradation and Delta‐like ligand 4 (DLL4) expression. These results indicate that VEGF‐A variants can be regulated by cell differentiation and are involved in IH progression. We also demonstrated that DLL4 expression was not exclusive to the endothelium in IH but was also present in pericytes, where the expression of VEGFR2 is absent, suggesting that pericyte‐derived DLL4 may prevent sprouting during involution, independently of VEGFR2. Angiogenesis in IH therefore appears to be controlled by DLL4 within the endothelium in a VEGF‐A isoform‐dependent manner, and in perivascular cells in a VEGF‐independent manner. The contribution of VEGF‐A isoforms to disease progression also indicates that IH may be associated with altered splicing. © 2016 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland.     

Angiogenesis and host immune response contribute to the aggressive character of non-melanoma skin cancers in renal transplant recipients

Mackenzie K A, Miller A P, Hock B D, Gardner J, Simcock J W, Roake J A, Dachs G U, Robinson B A & Currie M J
(2011) Histopathology  58 , 875–885
Angiogenesis and host immune response contribute to the aggressive character of non‐melanoma skin cancers in renal transplant recipients Aims: The aim of this study was to determine the contribution of tumour angiogenesis to the aggressive growth of non‐melanoma skin cancers (NMSCs) in renal transplant recipients (RTRs). Methods and results: The study cohort included RTRs (n = 38) with formalin‐fixed paraffin‐embedded tumour samples available from first post‐transplant NMSC (NMSC1) surgically excised at Christchurch Hospital, New Zealand, from 1997 to 2007. Comparable samples excised from immunocompetent individuals (ICIs) (n = 36) were selected to accommodate confounding factors. Markers of tumour angiogenesis were evaluated by immunohistochemistry, and analysed for associations with clinicopathological variables. As compared with ICIs, RTRs had a higher proportion of tumours with high microvessel density (P = 0.008), high proliferating capillary index (P < 0.0001) and low microvessel pericyte coverage index (P < 0.0001), and RTRs had a shorter cumulative second NMSC (NMSC2)‐free interval (P < 0.0001). ICIs had a higher proportion of tumours with a ‘marked’ number of vascular endothelial growth factor (VEGF)‐A‐positive leukocytes than RTRs (P = 0.04), and RTRs with a ‘moderate/marked’ number of VEGF‐A‐positive leukocytes had longer cumulative NMSC2‐free intervals than those with a ‘minimum’ number (P = 0.02). Conclusions: This study demonstrates increased tumour angiogenesis in NMSC in RTRs, and suggests a role for VEGF‐A‐positive peritumoural leukocytes in suppressing NMSC development.     

Time course of changes in angiogenesis-related factors in denervated muscle

Aim: Denervation leads to capillary regression in skeletal muscle. To gain insight into the regulation of this process, we investigated the time course of changes in capillary supply and gene expression of angiogenesis‐related factors during muscle denervation. Method: Female mice underwent surgery to transect the sciatic nerve, and then the gastrocnemius muscles were isolated at 12 h, 1, 3, 5, 10, 20, or 30 days after surgery. The capillary supply was assessed by immunohistochemistry using anti‐PECAM‐1/CD31 antibody. The mRNA levels for angiogenesis‐related factors were analysed using a real‐time polymerase chain reaction. Results: We found that the capillary‐to‐fibre ratio began to decrease 10 days after muscle denervation and decreased by 52% after 30 days. The levels of mRNA for vascular endothelial growth factor (VEGF), its receptors [fms‐like tyrosine kinase (Flt‐1) and a kinase insert domain‐containing receptor/fetal liver kinase‐1 (KDR/Flk‐1)], angiopoietin‐1 and angiopoietin‐2 of denervated muscle were immediately down‐regulated after 12 h and remained lower than control muscle until 30 days after muscle denervation. The levels of mRNA for the VEGF receptor, neuropilin‐1, angiopoietin receptor and Tie‐2 decreased within 12–24 h, but returned to near those of control muscle after 10–20 days, and again decreased after 30 days. Conclusions: These findings suggest that denervation‐induced capillary regression may be associated with down‐regulation of VEGF and angiopoietin signalling.     

Time‐course changes in VEGF expression and capillarity in the early stage of exercise training with Co2+ treatment in rat skeletal muscles

Aim: Cobalt administration was reported to mimic hypoxia. This study was designed to examine the time‐course changes in vascular endothelial growth factor (VEGF) expression and capillary geometry in skeletal muscles during endurance training with CoCl₂ administration in female Wistar rats. Methods: Exercise training by running lasted for up to 10 days at 25 m min^?1 on a 20% gradient, 15–42 min day^?1. Rats in the Co²⁺‐treated groups drank water containing 0.01% CoCl₂. Serial frozen sections were stained for alkaline phosphatase and dipeptidylpeptidase IV to identify capillary profiles and VEGF‐A protein. Results: In the soleus muscle, the density of VEGF‐positive capillaries (VEGF‐cap) was significantly increased after 6 and 10 days of the Co²⁺ administration (by 27 and 65% respectively) while the capillary‐to‐fibre ratio (C : F) first increased after 10 days. The training with Co²⁺ significantly increased VEGF‐cap by 69, 44 and 60%, respectively, after 3, 6 and 10 days. The VEGF‐cap was significantly increased after 6 and 10 days of training alone by 38 and 58%, respectively. In a similar extent, both training groups with and without Co²⁺ showed a significant increase in the C : F ratio after 6 and 10 days. Conclusions: The present results suggest that activation of the cellular oxygen‐sensing mechanism induced by Co²⁺ administration slightly facilitates an expression of VEGF but does not facilitate exercise‐induced microvascular remodelling in hind‐leg muscles.     

Overexpression of VEGF-A induces neovascularization and increased vascular leakage in rabbit eye after intravitreal adenoviral gene transfer

Aims: The aim of this study was to determine dose–response effects of vascular endothelial growth factor A as delivered using an adenoviral vector on vascular growth and pathological changes in the rabbit eye. Moreover, we wanted to develop a large animal model for angioproliferative diseases in the eye. Methods: Seventeen New Zealand White rabbits were injected with adenoviral vascular endothelial growth factor‐A (AdVEGF‐A) intravitreally with different doses (10⁹–10¹¹ vp). Controls were injected with an empty virus (AdCMV). Some animals had a combination of AdVEGF‐A and AdsKDR (a soluble form of the VEGF receptor‐2). Animals were killed 6 days after the gene transfer. On the basis of these results, 14 rabbits were injected intravitreally with AdVEGF‐A or adenoviral LacZ (AdLacZ) with 10¹⁰ vp in a volume of 0.1 mL. Animals were killed 3, 6, 14 and 28 days after the gene transfer, eyes were removed and analysed histologically. Results: In enzyme‐linked immunosorbent assay (ELISA) analysis, human VEGF‐A was present in vitreous humour in all VEGF‐A transduced eyes. The amount of VEGF‐A showed a dose‐dependent increase with the AdVEGF‐A dose and was the highest 6 days after the gene transfer. Histologic analyses revealed an increased capillary area and density in the AdVEGF‐A eyes when compared with the AdLacZ eyes (P < 0.05). In the AdVEGF‐A/AdsKDR eyes the average capillary area was not increased compared with AdLacZ eyes. Conclusion: This model could be useful for large animal studies regarding the pathogenesis of neoangiogenesis and for the development of new therapeutic strategies for angioproliferative diseases of the eye. Our results establish the key role of VEGF‐A in the induction of neovascularization and pathological changes in the rabbit eye.     

Lymphangiogenesis and Prognostic Significance of Vascular Endothelial Growth Factor C in Gastro‐oesophageal Junction Adenocarcinoma

Vascular endothelial growth factor C (VEGF‐C) is a crucial regulator of the development of lymphatic vessels and is involved in the lymph node metastasis of cancer. The levels of VEGF‐C expression and lymphatic vessel density (LVD) in 128 gastro‐oesophageal junction adenocarcinoma (GEJA) tissues were examined by immunohistochemistry and analysed for their association with clinicopathological features and disease‐free survival. We found that 75.0% of tumour samples displayed strong immunoreactivity to VEGF‐C. The levels of VEGF‐C expression in the tumour tissues were associated with the stages of the clinical tumours and the lymph node metastasis status, but not with the age, gender and the size and type of tumours in the cohort. Similarly, LVD, as evaluated by anti‐D2‐40 staining, was also associated with the clinical stages of GEJA. The values of LVD were positively correlated with the levels of VEGF‐C expression in these samples (r = 0.3760, P = 0.0001). High levels of VEGF‐C expression and high values of LVD were associated with shorter periods of disease‐free survival (DFS) in patients with GEJA (P < 0.001). In addition, GEJA at N1 and N2 stages, at T4 stage, chemotherapy after surgery, high levels of VEGF‐C expression and lower marginal resection were independent factors for the prognosis of DFS in patients with GEJA. Our data indicate that VEGF‐C may promote the lymphangiogenesis and lymphatic metastasis of GEJA and that VEGF‐C may be a valuable biomarker for the diagnosis of lymphatic metastasis and a prognostic factor of the survival of patients with GEJA.     

Differential expression of angiogenic factors in peripheral nerve sheath tumors

It is difficult to differentiate some malignant peripheral nerve sheath tumors (MPNST) from benign peripheral nerve sheath tumors (BPNST) histologically, and to predict the clinical outcome of patients with MPNST. In this study, the expression of VEGF and MVD were evaluated immunohistochemically in 22 cases of MPNST, 14 of neurofibroma and 19 of schwannoma and correlation of the staining grade of VEGF or MVD and the various clinical factors were analyzed, and statistically evaluated. Levels of VEGF mRNA expression were also determined with real-time RT-PCR. Statistically higher positive staining for VEGF was observed in MPNST compared to neurofibroma (P = 0.004) and schwannoma (P < 0.001). Even low grade MPNST showed higher VEGF positive staining than neurofibroma. Moreover, high VEGF expression statistically correlated with the poor prognosis of the patients with MPNST (P = 0.015). Although MVD in MPNST was significantly higher than that in neurofibroma (P = 0.038) and schwannoma (P < 0.001), MVD could not predict the prognosis of the patients with MPNST. Although VEGF mRNA expression tended to be higher in MPNST compared to neurofibroma, the difference was not significant. Levels of VEGF protein expression serve as a novel diagnostic and prognostic tools for peripheral nerve sheath tumors.     

VEGF/VEGFR2 and PDGF-B/PDGFR-β expression in non-metastatic renal cell carcinoma: a retrospective study in 1,091 consecutive patients

Purpose: We aimed to investigate the correlations between the expression of VEGF, PDGF-B, and their receptors (VEGFR2 and PDGFR-β) with pathologic stage or cell type in non-metastatic renal cell carcinoma. Materials and methods: VEGF, VEGFR2, PDGF-B, and PDGFR-β protein expression were evaluated immunohistochemically in prospectively collected 1,423 tumour samples obtained during radical or partial nephrectomy at a tertiary referral center. Intensity of expression was quantified on a scale of 0 to 3, and was compared among renal cell carcinoma cell types. Results: The study cohort consisted of 1,091 patients, of mean age 54 years, including 968 (88.7%) with clear cell, 82 (7.5%) with papillary, 31 (2.8%) with chromophobe, 4 (0.4%) with unclassified, and 6 (0.5%) with other types of renal cell carcinoma. VEGF expression increased with higher T and N stage and Fuhrman nuclear grade. PDGFR-β expression was highest in clear cell renal cell carcinoma, whereas VEGF and PDGF-B expression were highest in papillary renal cell carcinoma. After adjusting for T stage and Fuhrman nuclear grade using multivariate logistic regression analysis, VEGF (OR = 3.57, P < 0.001), VEGFR2 (OR = 1.82, P = 0.017), and PDGF-B (OR = 2.46, P = 0.019) expression were significantly greater in papillary than in clear cell type. Conclusions: Our results indicate that the cytoplasmic expression of VEGF, VEGFR2, PDGF-B, and PDGFR-β in RCC tumour cells is different in various pathologic stage and cell type. Notably, VEGF and PDGF-B expression are higher in papillary than in clear cell renal cell carcinoma. Further studies using quantitative measurement of proangiogenic factors in tumour cell are needed.     

Lymphangiogenesis in regional lymph nodes predicts nodal recurrence in pathological N0 squamous cell carcinoma of the tongue

Hirota K, Wakisaka N, Sawada‐Kitamura S, Kondo S, Endo K, Tsuji A, Murono S & Yoshizaki T
(2012) Histopathology
Lymphangiogenesis in regional lymph nodes predicts nodal recurrence in pathological N0 squamous cell carcinoma of the tongue Aims: Cancer cells induce de‐novo lymphatic vessel growth within draining lymph nodes before they metastasize. The aim of this study was to retrospectively evaluate lymph node lymphangiogenesis before the establishment of nodal recurrence in squamous cell carcinoma (SCC) of the tongue. Methods and results: Surgical specimens from 28 patients with pT2–T3N0M0 SCC of the tongue after local excision with supraomohyoid neck dissection were studied by immunohistochemistry. Intranodal lymphatic endothelium was highlighted by podoplanin staining to evaluate lymphatic vessel counts (LVCs). Primary tumour sections were examined for the expression of lymphangiogenic factors: vascular endothelial growth factor (VEGF)‐C and VEGF‐D. LVCs in regional lymph nodes were significantly increased in the cases with nodal recurrence (P = 0.0013). Simultaneous increases in VEGF‐C and VEGF‐D expression were significantly associated with both an increase in LVC in regional lymph nodes (P = 0.0001) and a decrease in the rate of survival without nodal recurrence (P = 0.016). Conclusions: Knowing the status of lymphangiogenesis in the regional pN0 lymph nodes in tongue cancer would help in predicting which patients will develop nodal recurrence. The use of a therapeutic approach which blocks lymphangiogenic factors, such as VEGF‐C and VEGF‐D, may be beneficial in suppressing the lymphatic spread of tongue cancer with intense intranodal lymphangiogenesis.     

血管内皮细胞生长因子及其受体2在慢性砷染毒大鼠睾丸中的表达及其意义*

目的：研究血管内皮细胞生长因子（ VEGF）及其受体2（ VEGFR2）在慢性砷中毒大鼠睾丸中的表达及意义。 方法：健康清洁级雄性SD大鼠随机分为高、中、低剂量砷染毒组和对照（ 蒸馏水）组,采用经口自由饮用方式 进行染毒,连续染毒6 个月。H-E 染色观察睾丸一般形态学变化,免疫荧光化学法对睾丸组织VEGF 和VEGFR2 的表达进行定位,采用实时荧光定量PCR法检测睾丸组织VEGF 和VEGFR2 的mRNA表达变化,流式细胞术观 察精子的凋亡率。结果：与对照组比较,中、高剂量砷组大鼠体质量明显降低,睾丸组织质量也明显降低,而低 剂量砷组大鼠体质量、睾丸质量、睾丸脏器系数差异无统计学意义。对照组睾丸结构正常,染砷组大鼠睾丸组织 上皮结构疏松,层次排列紊乱,层次逐渐减少,精原细胞出现空泡样改变,睾丸间质充血、渗出等,尤其中、高 剂量砷组有较明显的变化。与对照组比较,各染毒组VEGF 和VEGFR2 的mRNA表达水平均较低,砷染毒组大鼠 精子凋亡率较高,差异均有统计学意义。结论：慢性砷中毒时,VEGF 可能通过旁分泌- 自分泌的方式参与调控 大鼠睾丸的功能,VEGF 及其受体2 在砷中毒大鼠精子发生和发育过程中起重要作用。     

Vascular Endothelial Growth Factor Is Increased in Patients With Preeclampsia

PROBLEM: This study was conducted to determine whether altered levels of vascular endothelial growth factor (VEGF) may play a role in the pathogenesis of preeclampsia. METHOD OF STUDY: Maternal plasma samples were collected from 19 patients with preeclampsia (group A) either before the onset of labor, or before induction of labor or medical intervention. Plasma samples were also obtained from 19 normotensive patients with uncomplicated pregnancies (group B), who were matched with the patients with preeclampsia for gestational age and parity. Samples were frozen at ?70°C until assayed for VEGF by a specific enzyme-linked immunoassay. RESULTS: The mean maternal age was similar in groups A and B. For both groups the VEGF was detectable in all plasma samples. However, the plasma concentrations of VEGF were significantly increased in the group A patients, compared with those in group B (median, 47ng/ml; range, 10.6–72 ng/ml versus median, 13.6 ng/ml; range, 0.66-20 ng/ml; P < 0.001). In group A, a positive correlation was noted between VEGF concentrations and the systolic and diastolic blood pressure (r = 0.56; P = 0.01 and r = 0.48; P = 0.037, respectively). CONCLUSIONS: Maternal plasma VEGF levels were elevated in the patients with preeclampsia and correlated with the severity of hypertension, suggesting a role for VEGF in the pathogenesis of preeclampsia.     

Vascular endothelial growth factor A protein level and gene expression in intracranial meningiomas with brain edema

Meningiomas are the second most common primary intracranial tumors in adults. Although meningiomas are mostly benign, more than 50% of patients with meningioma develop peritumoral brain edema (PTBE), which may be fatal because of increased intracranial pressure. Vascular endothelial growth factor (VEGF) is an endothelial cell-specific mitogen and angiogen. VEGF-A protein, which is identical to vascular permeability factor, is a regulator of angiogenesis. In this study, 101 patients with meningiomas, and possible co-factors to PTBE, such as meningioma subtypes and tumor location, were examined. Forty-three patients had primary, solitary, supratentorial meningiomas with PTBE. In these, correlations in PTBE, edema index, VEGF-A protein, VEGF gene expression, capillary length, and tumor water content were investigated. DNA-branched hybridization was used for measuring VEGF gene expression in tissue homogenates prepared from frozen tissue samples. The method for VEGF-A analysis resembled an ELISA assay, but was based on chemiluminescence. The edema index was positively correlated to VEGF-A protein (p = 0.014) and VEGF gene expression (p < 0.05). The capillary length in the meningiomas was positively correlated to the PTBE (p = 0.038). If VEGF is responsible for the formation of PTBE, the edema may be treated with the anti-VEGF drug Bevacizumab (Avastin), which has been shown to reduce PTBE in patients with glioblastoma multiforme.     

Autologous cytokine‐induced killer (CIK) cell immunotherapy combined with cyclophosphamide in five patients with POEMS syndrome

The primary objective of this study was to evaluate the safety and clinical efficacy of autologous cytokine‐induced killer (CIK) cells combined with cyclophosphamide in the treatment of polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy and skin changes (POEMS) syndrome patients. We evaluated five POEMS syndrome patients treated with autologous CIK cell immunotherapy combined with cyclophosphamide from 1 May 2012 to 30 November 2014. The Overall Neuropathy Limitation Scale (ONLS), computed tomography of the chest and abdomen, ultrasound of the abdomen, serum vascular endothelial growth factor (VEGF) level and lymphocyte count findings in the five patients were recorded. The median age of the patients was 40 years (range: 25–62), and all the patients were male. CIK cells were generated routinely from peripheral blood mononuclear cells (PBMCs) of all five patients, and the numbers of CIK cells increased by approximately 105‐fold after 14 days of culture. All five patients (100%) responded to their neuropathy treatment, the ONLS scores were reduced by at least 1 and a paired‐sample t‐test revealed a significant difference (t = 5·715, P = 0·003 < 0·01). The extravascular volume overload responses indicated partial remission (PR = 60%) or stable disease (SD = 40%), and no cases of progressive disease (PD) or complete remission (CR) were observed. During clinical treatment, the serum VEGF of patient 5 decreased after one cycle of transfusion within 1 month. The lymphocyte counts of all the patients increased significantly after CIK transfusion, and a paired‐sample t‐test revealed a significant difference (t = 5·101, P = 0·004 < 0·01). Autologous CIK cell infusion combined with cyclophosphamide was found to be highly safe and elicited no adverse reactions. CIK cells can improve both the symptoms and quality of life, decrease serum VEGF levels and increase lymphocyte counts in patients with POEMS syndrome.     

Attenuation of the exercise‐induced increase in skeletal muscle Flt‐1 mRNA by nitric oxide synthase inhibition

We investigated the vascular endothelial growth factor (VEGF) receptor [fms‐like‐tyrosine kinase (Flt‐1 and fetal liver kinase‐1 (Flk‐1)] response to acute exercise. In female Wistar rats, the VEGF receptor messenger RNA (mRNA) response to a single acute exercise bout was examined using semi‐quantitative Northern blot from the left gastrocnemius muscles at rest and post‐exercise at 0, 1, 2, 4, 8, 16, 24 and 48 h. Exercise altered both Flt‐1 and Flk‐1 mRNA, with significant increases in Flt‐1 mRNA at 1 and 24 h. However, post‐hoc analysis was unable to discern the time point where a significant increase in Flk‐1 mRNA occurred. To investigate the regulation of Flt‐1 mRNA by exercise we examined if nitric oxide synthase (NOS) inhibition alters the Flt‐1 mRNA response. Eight groups [Condition: Rest or Exercise; Drug: Saline, 30 mg kg^–1N^ω‐nitro‐L ‐arginine methyl ester (L ‐NAME), 300 mg kg^–1L ‐NAME or 300 mg kg^–1D ‐NAME] were used to determine the effect of NOS inhibition on the Flt‐1 mRNA response to exercise. L ‐NAME, a known NOS inhibitor, attenuated the exercise‐induced increase in Flt‐1 mRNA by ～50%. These findings suggest that: (1) exercise alters Flt‐1 and Flk‐1 gene expression; and (2) NO is important in the regulation of the Flt‐1 gene response to exercise.     

血管内皮细胞生长因子在肥胖乳腺癌患者乳腺癌组织中的表达及临床意义

 摘要：　目的 探讨肥胖乳腺癌患者乳腺癌组织中血管内皮细胞生长因子（vascular endothelial growth factor,VEGF）的表达及临床意义。方法 采集45例肥胖乳腺癌患者,37例正常体重乳腺癌患者,用RT-PCR和免疫组织化学检测肥胖组和正常体重组乳腺癌组织VEGF的mRNA和蛋白表达。结果 肥胖组乳腺癌组织VEGF的mRNA和蛋白表达均高于正常体重组 (P<0.05),并且VEGF mRNA和蛋白表达之间呈正相关(r=0.785,P<0.05);VEGF在肥胖乳腺癌组织中的表达与组织学分级、5年复发转移有关,与患者年龄无关。结论 VEGF在肥胖乳腺癌患者中具有高表达趋势,提示VEGF可能在肥胖相关性乳腺癌的发生、演变及预后中起重要作用。