Clinical diagnosis and prognostic value of serum TLR4 and VEGFA expression in diabetic retinopathy / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To investigate the expression and clinical significance of Toll-like receptor 4(TLR4)and vascular endothelial growth factor A(VEGFA)in the serum of patients with diabetic retinopathy(DR).METHODS: A total of 183 patients with type 2 diabetes mellitus(T2DM)admitted to our hospital from January 2021 to January 2022 were collected as the study subjects. They were grouped into non diabetic retinopathy(NDR)group(n=54), proliferative diabetic retinopathy(PDR)group(n=68)and non proliferative diabetic retinopathy(NPDR)group(n=61). In the same period, 70 volunteers who underwent physical examination in our hospital were randomly stratified according to age and sex. After discharge, DR patients were followed up for 1a and grouped into a poor prognosis group(n=40)and a good prognosis group(n=89)based on whether they had visual impairment. Enzyme-linked immunosorbent assay(ELISA)was applied to detect the levels of TLR4 and VEGFA in serum; Logistic regression was applied to analyze the influencing factors of DR; receiver operating characteristic(ROC)curve was applied to analyze the clinical value of serum TLR4 and VEGFA levels in diagnosing DR and predicting prognosis.RESULTS: There were statistical significance in TLR4 and VEGFA levels among the control group, NDR group, PDR group, and NPDR group(F=935.753, 516.936, all P<0.05), and further pairwise comparisons showed statistical significance(P<0.05); the expression levels of TLR4 and VEGFA in the serum of patients with poor prognosis were higher than those of patients with good prognosis(P<0.01); the results of Logistic regression analysis showed that TLR4, VEGFA, course of disease, and HbA1c were all risk factors for the occurrence of DR(P<0.05); the ROC results showed that the AUC of serum TLR4, VEGFA levels, and their combination for predicting DR was 0.869, 0.862, and 0.931, respectively, the AUC of serum TLR4, VEGFA levels, and their combined prediction of visual disability in DR patients was 0.864, 0.863, and 0.938, respectively.CONCLUSION: The expression of TLR4 and VEGFA in serum of DR patients is up-regulated, and the combined detection of TLR4 and VEGFA can be used as a potential indicator to evaluate the occurrence and poor prognosis of DR.

Effect of Chaihu Guizhitang on Triple-negative Breast Cancer Cells： Based on HIF-1α/VEGFA Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the effect of Chaihu Guizhitang on triple-negative breast cancer （TNBC） cells based on hypoxia-inducible factor-1α （HIF-1α）/vascular endothelial growth factor A （VEGFA） signaling pathway. MethodTNBC xenograft model was established and the cells were randomized into model group， capecitabine group （0.2 mg·kg-1）， Chaihu Guizhitang low-dose group， medium-dose group， and high-dose group （10.62， 21.23， 42.46 g·kg-1）， with 10 mice in each group. After 21 days of medication， the content of tumor necrosis factor-α （TNF-α） in serum was detected by enzyme-linked immunosorbent assay （ELISA）. The expression of HIF-1α mRNA was detected by real-time fluorogenic quantitative polymerase chain reaction （real-time PCR）. Immunohistochemistry （IHC） was employed to detect the expression of HIF-1α， TNF-α， and VEGFA in tumor tissues， and CD34 staining to examine the angiogenesis in tumor tissues. Microvessel density （MVD） was calculated， and the protein expression of HIF-1α， VEGFA， and epidermal growth factor receptor （EGFR） in tumor tissues was measured by Western blot. ResultCompared with the model group， the rest four groups showed low levels of TNF-α （P<0.01）， HIF-1α mRNA （P<0.01）， expression of HIF-1α， TNF-α， VEGFA， and CD34 in cells， and MVD （P<0.05， P<0.01）， and low protein levels of HIF-1α， VEGFA， and EGFR （P<0.01）. Compared with capecitabine group， medium-dose and high-dose Chaihu Guizhitang decreased the level of TNF-α （P<0.01）， HIF-1α mRNA （P<0.01）， expression of HIF-1α， TNF-α， and VEGFA in cells （P<0.01）， CD34 expression， MVD， and protein levels of HIF-1α， VEGFA， and EGFR （P<0.01）. ConclusionChaihu Guizhitang may inhibit the angiogenesis in TNBC cells by regulating the expression of HIF-1α/VEGFA signaling pathway， thus exerting anti-tumor effect.

Mechanism of Astragaloside Ⅳ on HepG2 Cells Based on Molecular Dynamics Simulation and Experimental Evaluation / 肿瘤防治研究

Objective To reveal the mechanism of action of AS-Ⅳ on HepG2 cells based on molecular dynamics simulation and experimental evaluation. Methods We constructed a "drug-disease" network pharmacological map, analyzed the core genes of astragaloside Ⅳ (AS-Ⅳ) in HCC, screened key signaling pathways, and established a "drug-target" molecular dynamics model. In vitro assay was used to detect migration, proliferation and invasion abilities. Flow cytometry and qRT-PCR were used to detect the effect of AS-Ⅳ on the cell cycle and apoptosis, and the expression of core gene of HepG2. Results The core target of AS-Ⅳ acting on HCC was VEGFA. Compared with the control group, the high concentration of AS-Ⅳ significantly inhibited the migration, invasion and proliferation of HepG2 cells, blocked the metastasis of HepG2 cells from G1 to G2 phase, promoted their apoptosis, down-regulated VEGFA expression and up-regulated TGF-β1 expression. Conclusion AS-Ⅳ may inhibit the proliferation of hepatocellular carcinoma cells through multi-target and multi-pathway.

Regulatory Mechanism of Zedoary Turmeric Oil on VEGFA， STAT3 and mTOR in Ovarian Cancer / 中国实验方剂学杂志

Objective:To explore the effects and mechanism of zedoary turmeric oil and its active components on the vascular endothelial growth factor A （VEGFA）， signal transducer and activator of transcription 3 （STAT3）， and mechanistic target of rapamycin （mTOR） in the ovarian cancer （OC）. Method:Network pharmacology technology was employed to analyze the mechanism of Curcumae Rhizoma on OC. Bioinformatics was used to analyze the expression of VEGFA， STAT3， and mTOR in OC and the effect on the prognosis of OC to explore the feasibility of zedoary turmeric oil in regulating VEGFA， STAT3， and mTOR in OC.The xenograft tumor model of nude mice was established， and the effects of zedoary turmeric oil and its active components on VEGFA， STAT3， and mTOR in OC were observed by hematoxylin-eosin （HE） staining， real-time fluorescence-based quantitative polymerase chain reaction （Real-time PCR）， Western blot， and immunohistochemistry （IHC）. Result:Bioinformatics analysis and literature research showed that VEGFA， STAT3， and mTOR played a special regulatory role in the occurrence and development of OC， and were potential key targets for the proliferation of OC. Network pharmacology analysis revealed that Curcumae Rhizoma could regulate multiple disease targets of OC， and mediate VEGFA， STAT3， and mTOR in OC through these multiple targets. As demonstrated by HE staining， the tumor cells in the model group were densely arranged， with no erosion on the edge and no vesicles inside. Compared with the model group， the cell density in other treatment groups was reduced， and strip-shaped erosion on the edge and small empty vesicles were observed in the tumor tissue， especially in the zedoary turmeric oil group. According to the results of Real-time PCR and IHC， zedoary turmeric oil and its active components could inhibit the mRNA and protein expression of VEGFA， STAT3， and mTOR in the OC tissue （<italic>P</italic><0.05）. Conclusion:Zedoary turmeric oil and its active components could reduce the expression of VEGFA， STAT3， and mTOR in tumor tissue of nude mice， and inhibited the proliferation of OC through VEGFA， STAT3， and mTOR.

Experimental study of 125I seeds inhibiting angiogenesis of transplanted hepatocellular carcinoma / 中华放射医学与防护杂志

Objective:To investigate the effect of radioactive 125I seed on angiogenesis of subcutaneously transplanted hepatocellular carcinoma in nude mice and underlying mechanism. Methods:The subcutaneous transplanted tumor model of human hepatocellular carcinoma Huh7 cells was established in nude mice. Twelve nude mice were randomly divided into observation group and control group with 6 mice in each group. The 125I seed with activity of 2.96×10 7Bq was implanted into the transplanted tumor of observation group and another with 0 Bq as control group, respectively. The volume of the transplanted tumor was measured every 4 d and the growth curve of the tumor was recorded. The microvessel density (MVD) of the transplanted tumor was evaluated by immunohistochemical detection of CD31. VEGF-A and HIF-1α protein and mRNA were detected by immunohistochemistry and RT-PCR, respectively. Results:The growth rate of tumor in the observation group was slower than that in the control group, and the difference of tumor volume between two groups at 12 d after 125I seed implantation was significantly different( t=3.167, P<0.05). At 28 d after transplantation, the tumor volumes of control and observation group approached to (963.61 ± 89.56) mm 3and (602.10±75.98) mm 3, respectively. The MVD of the observation group was significantly lower than that of the control group ( t=6.361, P<0.05). The relative expression of VEGF-A and HIF-1α mRNA in the observation group was significantly lower than that in the control group ( t=10.480, 6.414, P<0.05). Protein expression levels of VEGF-A and HIF-1α in the observation group were lower than those in the control group ( t=10.890, 12.250, P<0.05). Conclusions:Radioactive 125I seed can inhibit the growth of HCC xenografts by reducing tumor microvessels, which may be related to the decrease of VEGF-A and HIF-1α expression.

miR-17-5p mitigates endometriosis by directly regulating VEGFA

Endometriosis is a common disease in women, which impairs the quality of life in patients. Recently, accumulating evidences reported that miRNAs play an essential role in diagnosis and treatment of endometriosis.However, the regulatory mechanism of miRNAs has not been fully explored. The expression of miR-17-5p andVEGFA was detected using qRT-PCR. The protein level of VEGFA was measured via Western blot. Cellproliferation was determined by CCK-8 assay. Cell migration and invasion were measured via transwell assay.The relationship of miR-17-5p and VEGFA were verified via luciferase reporter assay. Then miR-17-5p wasremarkably down-regulated in endometriosis tissues, serums and cells, and overexpression of miR-17-5pinhibited cell proliferation, migration and invasion in endometriosis. Results showed that VEGFA was significantly up-regulated in endometriosis tissues and cells and acted as a target of miR-17-5p. Moreover, miR17-5p negatively regulated VEGFA expression in endometriosis. Otherwise, up-regulation of VEGFAimproved cell proliferative, migrated and invasive ability in ECSCs with transfection of miR-17-5p mimicsgroup. Our data showed miR-17-5p inhibits cell proliferation, migration and invasion in endometriosis bydirectly repressing VEGFA expression.

Diagnostic value of combined detection of multiple serum cytokines for gastric adenocarcinoma / 中国肿瘤生物治疗杂志

@#[Abstract] Objective:To explore the clinical significance of multiple serumcytokines in early diagnosis and progression assessment of gastric adenocarcinoma. Methods: Peripheral blood samples of 85 healthy subjects (healthy control group) and 81 patients with pathologically confirmed gastric adenocarcinoma (gastric cancer group) were collected from November 2017 to February 2018 at Shanxi Cancer Hospital. Serum levels of 17 cytokines (including IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p70, IL-15, IL-17A, TNF-α, TNF-β, GM-CSF, G-CSF, IFN-γ, IP-10, MCP-1 andVEGF-A) were measured byAimPlex multiplex assay technology.Their diagnostic values were analyzed by receiver operating characteristic (ROC) curve. Results: Serum levels of IL-10, IL-8, IL-6, IP-10, MCP-1, VEGF-Aand IL-12p70 were significantly higher in gastric cancer patients than those in healthy controls (all P<0.01). There were significantly increasedlevelsofIL-8,IL-6and VEGF-Ain advanced-stage gastriccancer(stageI/II)groupoverearly-stage gastric cancer (stage III/IV) group (all P<0.01).AUC (areas under the curve) of IL-8, IL-6, IL-10, IP-10, MCP-1, IL-12p70 and VEGF-Afor distinguishing early-stage gastric cancer patientsfromhealthy controls was0.98,0.92,0.89,0.84,0.76,0.74 and 0.58, respectively. The diagnostic sensitivity of IL-8, IL-6 and IL-10 was 97.4%, 89.5% and 97.4%, respectively, and the specificity was 87.1%, 85.9%and 77.6%, respectively.TheAUCof IL-8, IL-6 andVEGF-Afor distinguishing advanced-stage gastric cancer patients from early-stage gastric cancer patients was 0.82, 0.72 and 0.69, respectively. Thediagnosticsensitivity of IL-8, IL-6 and VEGF-A was 83.7%, 60.5% and 41.9%, respectively, and the specificity was71.1%,76.3%and 92.1%, respectively. Conclusion: ThecombineddetectionofserumIL-8,IL-6andIL-10 may be a potential approach for early screening of gastric adenocarcinoma, which canalsobeusedtoassessthe progression of gastric adenocarcinoma.

Effect of triptolide in improving platelet activation in patients with ankylosing spondylitis by regulating VEGFA,SDF-1,CXCR4 pathway / 中国中药杂志

The effect of triptolide( TP) on VEGFA,SDF-1,CXCR4 pathway were investigated in vitro to explore the mechanism in improving platelet activation in patients with ankylosing spondylitis( AS). Peripheral blood mononuclear cells( PBMC) were used for the experiment and divided into 4 groups: normal group( NC),model group( MC),triptolide group( TP),and AMD3100 group. The optimal concentration of TP was measured by the MTT method. The expressions of TNF-α,IL-1β,IL-4,IL-10,VEGFA and VEGFR were detected by ELISA. The expressions of SDF-1,CXCR4 and VEGFA were detected by real-time quantitative PCR( RT-qPCR).The expressions of SDF-1,CXCR4,VEGFA and VEGFR were detected by Western blot. The expression levels of CD62 p,CD40 L and PDGFA were detected by immunofluorescence. MTT results showed that medium-dose TP had the strongest inhibitory effect on cells at24 h. The results of ELISA and PCR showed that TP inhibited mRNA expressions of IL-1β,TNF-α,VEGFA,VEGFR and SDF-1,CXCR4 and VEGFA. The results of Western blot indicated that TP inhibited SDF-1,CXCR4 and VEGFA,VEGFR protein expressions; immunofluorescence results indicate that TP can inhibit the expressions of CD62 p,CD40 L,PDGFA. TP may regulate platelet activation by down-regulating SDF-1,CXCR4,VEGFA and VEGFR mRNA expressions,thereby down-regulating IL-1β and TNF-αexpressions,and up-regulating the expressions of IL-4 and IL-10 cytokines.

lncRNA SBF2-AS1 promotes epithelial-mesenchymal transition of cervical cancer cells via regulating miR-140-5p/VEGFAaxis / 中国肿瘤生物治疗杂志

@# Objective: To investigate the effect of lncRNA SBF2-AS1 on epithelial-mesenchymal transition (EMT) of cervical cancer HeLa cell via regulating miR-140-5p/VEGFA (vascular endothelial growth factor A) axis. Methods: After cell culture and transfection, the cells were divided into 5 groups: NC group, miR-140-5p mimic group, miR-140-5p mimic+pcDNA-VEGFA group, si-lncRNA SBF2-AS1+pcDNA-VEGFA group and si-lncRNA SBF2-AS1+miR-140-5p mimic group. The expression level of lncRNA SBF2-AS1 in cervical cancer tissues and cell lines was detected by qPCR. The targeted relationship between lncRNA SBF2-AS1, miR-140-5p and VEGFA was confirmed by Dual luciferase reporter gene assay. The expression levels of VEGFA and EMT-related proteins N-cadherin, Vimentin and E-cadherin in HeLa cells were detected by WB. The invasion and migration of HeLa cells were detected by Transwell. Results: lncRNA SBF2-AS1 was highly expressed in cervical cancer tissues and cell lines (P<0.05 or P<0.01). Dual luciferase reporter gene assay confirmed that lncRNASBF2-AS1 targetedly combined with miR-140-5p and VEGFAwas a target gene of miR-140-5p (P< 0.05). Knockdown of lncRNA SBF2-AS1 inhibited invasion and migration as well as EMT of HeLa cells. Further experiment confirmed that lncRNA SBF2-AS1 up-regulated the expression level of VEGFA via miR-140-5p, thereby promoting invasion, migration and EMT of HeLa cells. Conclusion: lncRNASBF2-AS1 promotes EMT of HeLa cells via miR-140-5p/VEGFAaxis.

Therapeutic effect of total triterpenoids of Chaenomeles speciosa combined with omeprazole on gastric ulcer induced by indomethacin in rats / 中国中药杂志

The aim of this paper was to observe the combination therapy with total triterpenoids of Chaenomeles speciosa and omeprazole on indomethacin-induced gastric ulcer in rats, and explore its possible mechanism. Rats were randomly divided into normal group, model group, omeprazole monotherapy(3.6 mg·kg~(-1)) group, total triterpenoids of C. speciosa monotherapy(100 mg·kg~(-1)) group, total triterpenoids of C. speciosa and omeprazole combination therapy(100 mg·kg~(-1)+3.6 mg·kg~(-1)) group. Except for the normal group, the other groups were given indomethacin(20 mg·kg~(-1)) by oral once a day for 7 consecutive days. Then the treated groups were given corresponding drugs by gavage, once a day for 14 consecutive days. The next day after the last administration, half of the rats in each group were measured the gastric mucosal blood flow, gastric juice volume and serum TNF-α, IL-1β, IL-6, IL-4 and IL-10. After the remaining rats in each group were underwent pyloric ligation 4 hours after the last administration, the gastric endocrine volume, pH value and total acidity of gastric secretion were measured, then histological analysis was performed, MPO activity, cAMP content and histomorphological analysis were conducted. Real-time PCR was applied to detect the mRNA expressions of gastric tissue TNF-α,IL-1β, IL-6, IL-4, IL-10, VEGFA, A_(2A)R; the protein expressions of VEGFA, A_(2A)R, PKA, p-PKA, CREB, p-CREB, EGF, EGFR, p-EGFR, MUC6, TFF2 in gastric tissue were detected by Western blot. The results indicated that total triterpenoids of C. speciosa and omeprazole combination therapy might significantly increase gastric mucosal blood flow, gastric mucus volume, reduce gastric endocrine volume, secretion acidity and mucosal damage, decrease the levels of TNF-α,IL-1β and IL-6, increase the levels of IL-4 and IL-10 in blood and gastric tissue, inhibit the activity of MPO, increase the content of cAMP in gastric tissue, up-regulate the mRNA expressions of VEGFA, A_(2A)R and protein expressions of VEGFA, A_(2A)R, PKA, p-PKA, CREB, p-CREB, EGF, EGFR, p-EGFR, MUC6, TFF2 in gastric tissue, elevate p-PKA/PKA, p-CREB/CREB and p-EFGR/EFGR. Moreover, the combination therapy with total triterpenoids of C. speciosa and omeprazole was more obvious than those of two monotherapies. These aforementioned findings suggested that the combination therapy with total triterpenoids of C. speciosa and omeprazole on indomethacin-induced gastric ulcer have significant therapeutic effect on indomethacin induced gastric ulcer in rats, its mechanism might be related to regulating A_(2A)R/AKT/CREB, A_(2A)R/VEGFA, EGF/EGFR and MUC6/TFF2 signaling pathways, inhibiting pro-inflammatory factors, increasing gastric mucosal blood flow, up-regulating mucosal cell proliferation factors and promoting mucosal protective factors.

Estrogen deficiency in ovariectomized rats: can resistance training re-establish angiogenesis in visceral adipose tissue?

OBJECTIVE: The purpose of this study was to investigate the effects of resistance training on angiogenesis markers of visceral adipose tissue in ovariectomized rats. METHOD: Adult Sprague-Dawley female rats were divided into four groups (n=6 per group): sham-sedentary, ovariectomized sedentary, sham-resistance training and ovariectomized resistance training. The rats were allowed to climb a 1.1-m vertical ladder with weights attached to their tails and the weights were progressively increased. Sessions were performed three times per week for 10 weeks. Visceral adipose tissue angiogenesis and morphology were analyzed by histology. VEGF-A mRNA and protein levels were analyzed by real-time PCR and ELISA, respectively. RESULTS: Ovariectomy resulted in higher body mass (p=0.0003), adipocyte hypertrophy (p=0.0003), decreased VEGF-A mRNA (p=0.0004) and protein levels (p=0.0009), and decreased micro-vascular density (p=0.0181) in the visceral adipose tissue of the rats. Resistance training for 10 weeks was not able to attenuate the reduced angiogenesis in the visceral adipose tissue of the ovariectomized rats. CONCLUSION: Our findings indicate that the resistance training program used in this study could not ameliorate low angiogenesis in the visceral adipose tissue of ovariectomized rats.

Reduced Vascular Endothelial Growth Factor A Levels are Correlated with the Incidence of Brain Edema in Acute Ischemic Stroke Patients.

Aims: To study the correlation between the vascular endothelial growth factor A/VEGF-A level and the incidence of brain edema in acute ischemic stroke patients. Study Design: A prospective observational analytic case-control study. Place and Duration of Study: Stroke Unit at the Dr. Sardjito General Hospital, Yogyakarta, Indonesia, between December 2010 and August 2011. Methodology: Seventy-one hospitalized acute ischemic stroke patients were recruited, consisting of 37 subjects in the brain edema group and 34 subjects in the non-brain edema group. Comparative analysis of the VEGF-A levels in blood was performed between the brain edema and non-brain edema groups. Results: The average level of VEGF-A in the brain edema group was 436 pg/mL and the one in the non-brain edema group was 746 pg/mL. This difference was statistically significant (95%CI: 5.5-615; P=.046). The proportion of VEGF-A levels less than the calculated cut-off point (638.3 pg/mL) in the brain edema group were significantly greater than the ones in the non-brain edema (83.78% and 58.82%, respectively; OR=3.6; 95%CI=1.06-13.26; P=.020). Conclusions: The decreased levels of VEGF-A in blood were correlated with the incidence of brain edema in acute ischemic stroke patients.

The Interaction of Stroke Risk Factors, Clinical Outcomes and Matrix Metalloproteinase 9 Levels with the Reduction of Vascular Endothelial Growth Factor A Levels at the Incidence of Brain Edema in Acute Ischemic Stroke Patients.

Aims: To study the interaction between the reduction of vascular endothelial growth factor A/VEGF-A level with other variables at the incidence of brain edema in acute ischemic stroke patients. Study Design: A prospective observational analytic case-control study. Place and Duration of Study: Stroke Unit at the Dr. Sardjito General Hospital, Yogyakarta, Indonesia, between December 2010 and August 2011. Methodology: Hospitalized acute ischemic stroke patients were recruited, comprising of 37 subjects in the brain edema group and 34 subjects in the non-brain edema group. Clinical characteristics of each subject were recorded and blood levels of VEGF-A and matrix metalloproteinase 9/MMP-9 were measured. Logistic regression analyses were performed to discover any potential independent variable that can influence the VEGF-A role at the incidence of brain edema. Results: Multivariate analyses revealed that several variables were significantly interacted with the reduction of VEGF-A levels at the incidence of brain edema. These variables were the lipid profiles (model “D”; OR=4.26; 95%CI: 1.28-14.15), stroke risk factors (model “G”; OR=4.78; 95%CI: 1.38-16.56), MMP-9 (model “I”; OR=5.59; 95%CI: 1.58-19.78), and Gadjah Mada Stroke Scale/GMSS score (model “K”; OR=5.29; 95%CI: 1.47-19.08). Subsequent multivariate analysis by combining the model “D”, “G”, “I” and “K” resulted in a very elevated odds ratio (OR=16.72; 95%CI: 2.75-101.5). Conclusion: The influence of VEGF-A reduction at the incidence of brain edema would be strongly enhanced by considering the combination of several independent variables, including the lipid profile, history of previous stroke risk factors, MMP-9 levels and GMSS score.

Genetic Risk Factors for Psoriasis in Turkish Population: -1540 C/A, -1512 Ins18, and +405 C/G Polymorphisms within the Vascular Endothelial Growth Factor Gene

BACKGROUND: Evidence regarding the vascular endothelial growth factor A (VEGFA) as a potent mediator of angiogenesis and inflammation in psoriasis has revealed variations in this gene as surrogate markers of psoriasis. OBJECTIVE: VEGFA gene polymorphisms (-1540 C/A, -1512 Ins18, -460 T/C, and +405 C/G) in psoriasis susceptibility in Turkish population were investigated. METHODS: A total of 200 age, sex and ethnicity-matched psoriatic and healthy individuals were examined for clinical type, response to therapy, serum VEGFA and its receptor levels, genotypes and haplotypes. RESULTS: The +405 GG, +405 CG, -1540 CA, and -1512 +Ins18 genotypes conferred a significant risk for developing psoriasis. The C-InsTC haplotype in the controls and C+InsTG, A+InsTC, and A-InsTG haplotypes in psoriatic patients were observed to be significantly high. Increased serum levels of VEGFA were detected in psoriatic patients with the C-InsTC haplotype than that in the controls. The +405 GG genotype was significantly more frequent in psoriatic patients with a positive family history, and the moderate form of psoriasis was more frequent among C+InsTG haplotype carriers than that among the other patients. The +405 GG genotype was found to be more frequent in patients responding to oral retinoids. Serum VEGFR1/FLT1 and VEGFR2/KDR levels were not significantly different when psoriatic patients and controls were stratified based on the risk polymorphic variants. CONCLUSION: VEGFA gene +405 GG and CG, -1512+Ins18, and -1540 CA genotypes are associated with an increased risk of psoriasis in Turkish population. The G allele at +405 and an 18-bp insertion at -1512 are primarily the risk factors for psoriasis, and this risk is potentiated by the presence of the A allele at the -1540 locus.

Effect of rosiglitazone on the expression of AQP-1, VEGF-A and COX-2 in uremic rat of peritoneal dialysis / 中华肾脏病杂志

Objective To investigate the effect of rosiglitazone(RGZ) on peritoneal morphology,function and the expressions of Aquaporin 1 (AQP-1),vascular endothelial growth factor A (VEGF-A) and cyclooxygenase 2(COX-2) in uremic rat of peritoneal dialysis.Methods Thirty Sprague-Dawley rats were randomly divided into five groups.Group S (n=6) was subjected to sham operation.Group N (n=6) was subjected to nephrectomy with silicon catheter inserted,but no peritoneal exposure.Group P (n=6) was subjected to nephrectomy with silicon catheter inserted and receiving daily peritoneal injection through the catheter,using 4.25％ peritoneal dialysis fluid 10 ml twice a day for 2 weeks.Group R (n=6) was subjected to nephrectomy with silicon catheter inserted and receiving daily peritoneal injection through the catheter,using 4.25％ peritoneal dialysis fluid containing rosiglitazone (0.2 mg/kg) 10 ml twice a day for 2 weeks.Group GW (n=6) was subjected to nephrectomy with silicon catheter inserted and receiving daily peritoneal injection through the catheter,using 4.25％ peritoneal dialysis fluid containing rosiglitazone (0.2 mg/kg) and GW9662 (0.2 mg/kg) 10 ml twice a day for 2 weeks.After two weeks of dialysis,a 90 min peritoneal equilibration test was performed and the amount of ultrafiltration was accurately measured.The partial peritoneum tissues of rats were harvested and stained by hematoxylin-eosin (HE),then morphology changes of partial peritoneum were examined by light microscopy.The expression of AQP-1,VEGF-A and COX-2 in omentum were detected with immunohistochemistry assay.AQP-1,VEGF-A and COX-2 mRNA were detected by qRT-PCR.Results Morphology changes of partial peritoneum showed that compared with Group S,a dramatic increase in thickness of the mesothelium-to-muscle layer of peritoneum in Group N,P,R and GW(P ＜0.05).Compared with group P,the thickness significantly decreased in Group R(P ＜ 0.05).PET results showed that compared with Group S,ultrafiltration (UF) significantly reduced in Group P,R,and GW (P ＜ 0.05).Compared with Group P,ultrafiltration significantly increased in Group P,R,and GW (P ＜0.05).Compared with group S,the expressions of AQP1,VEGF-A and COX-2 mRNA and protein were significantly increased in group P,R and GW(P ＜ 0.05).Compared with group P,the expressions of AQP1,VEGF-A mRNA and protein were significantly decreased in Group R and GW(P ＜ 0.05).Compared with group P,the expressions of COX-2 mRNA and protein were significantly decreased in group R (P ＜ 0.05),while no differences in the expression of COX-2 mRNA and protein in group GW (P ＜ 0.05).Conclusions Rosiglitazone can inhibit peritoneal interstitial and vascular proliferation,protect peritoneal function and increase ultrafiltration.Rosiglitazone can protect peritoneal function probably by inhibiting expression of VEGF-A and COX-2.

Study on correlation between VEGF level and malignant pleural effusion / 中国生化药物杂志

Objective Methods To explore the value of VEGF levels in serum and pleural effusion for diagnosis of benign and malignant tumors, and evaluate clinical value of VEGF-A, C, D in malignant pleural effusion.Methods Serum and pleural effusion of 34 cases patients with lung cancer were collected in our hospital, the application of ELISA method for the detection of VEGF level in serum and pleural effusion in patients with lung cancer and with benign pleural effusion.VEGF-A, C, D levels were detected.Results VEGF levels in serum and pleural effusion in malignant group were significantly higher than those in benign group(P<0.05).In addition, patients with lung cancer before initial treatment, the VEGF levels in serum and pleural effusion of distant metastasis group were significantly higher than those of without distant metastasis group(P<0.05).There was a correlation between the level of VEGF and the malignant pleural effusion(r=0.878, P<0.05).No correlation existed between VEGF level and benign pleural effusion.The content of sVEGF-A in serum had no statistical difference in cancer group and benign group.Effusion supernatants of pVEGF-A content in lung cancer group were higher than those in benign effusion group(P<0.05).pVEGF-A and sVEGF-A levels were similar in benign effusion group. Effusion supernatants pVEGF-A in malignant group was higher than that in benign effusion group(P<0.05).pVEGF-A was significantly higher than that of sVEGF-A in malignant effusion(P<0.05).Serum VEGF-C, VEGF-D content had no significant difference between cancer group and benign group. pVEGF-C, pVEGF-D content had no significant difference between cancer group and benign group.Conclusion Level of VEGF in serum and pleural effusion detection would help to diagnose and differentially diagnose benign and malignant pleural effusion.Effusion VEGF-A is different in benign and malignant effusion, which may become benign and malignant effusion tumor markers.

VEGFA SNPs (rs34357231 & rs35569394), Transcriptional Factor Binding Sites and Human Disease.

Purpose: The human vascular endothelial growth factor (VEGF)-A gene transcribes a signaling protein involved in the regulation of angiogenesis, vasculogenesis and endothelial cell growth. Two insertion/deletion (I/D) simple nucleotide polymorphisms (SNPs, rs34357231 & rs35569394) in the promoter region of the gene have been significantly associated with several human diseases. These SNPs were computationally examined with respect to changes in punitive transcriptional factor binding sites (TFBS) and these changes were discussed in relation to the diseases. Methods: The JASPAR CORE and ConSite databases were instrumental in identifying the TFBS. The Vector NTI Advance 11.5 computer program was employed in locating all the TFBS in the VEGFA gene from 2.7 kb upstream of the transcriptional start site to 1.6 kb past the 3’UTR. The JASPAR CORE database was also involved in computing each nucleotide occurrence (%) within the TFBS. Results: Regulatory SNPs (rSNPs) in the promoter region of the VEGFA gene alter the DNA landscape for potential transcriptional factors (TFs) to attach resulting in changes in TFBS. The VEGFA-deletion (D) allele of these SNPs has been found to be a risk factor for diabetic retinopathy, diabetic microvascular complications in patients with type 1 diabetes mellitus, breast cancer in north Indian patients, and bladder cancer. The changes in TFs associated with the TFBS are examined with respect to these human diseases. Conclusion: The VEGFA-insertion (I) allele provides punitive TFBS for the AR, EGR1 & 2, KLF5 and SP1 TFs whose BS do not occur with the VEGFA-D allele. These TFs have been linked to prostate cancer, cancer suppression and oncogenic processes and if not regulating the VEGFA gene may pose a risk for disease.

Effect of M-CSF and GM-CSF on migration and VEGF-A expression of breast cancer cell line 4T1 / 肿瘤研究与临床

Objective To investigate the effect of M-CSF and GM-CSF on migration and expression of VEGF-A in breast cancer cell line 4T1.Methods Real-time PCR was used to detect VEGF-A mRNA expression in 4T1 cells treated by 5 ng/ml or 10 ng/ml M-CSF or GM-CSF.Ability of migration and metastasis of 4T1 cells were analyzed by scratch and Transwell assays.Results The relative expression of VEGF-A mRNA at 12 h and 24 h in 4T1 cells treated by 5 ng/ml or 10 ng/ml M-CSF were 17.81±2.49 and 17.48± 5.43,5.15±2.59 and 5.45±4.28,respectively,while those treated by GM-CSF were 9.77±2.39 and 7.61±2.80,6.53±2.41 and 6.30±2.89,respectively.M-CSF and GM-CSF can promote the expression of VEGF-A in 4T1 cells (P ＜ 0.05).The relative expression of VEGF-A was higher in 4T1 cells treated for 12 h than that for 24 h (P ＜ 0.01).M-CSF,GM-CSF and VEGF-A can promote metastasis of 4T1 cells (all P ＜ 0.05),whereas no gross migration of 4T1 cells was showed by VEGF-A treatment.Conclusion M-CSF and GM-CSF can promote the migration and expression of VEGF-A in breast cancer cell line 4T1.

Radioresistance change and the mechanism of human esophaged cancer EC9706 cells in hypoxia / 中华放射医学与防护杂志

Objective To investigate the expression of hypoxia-inducible factor-1 alpha (HIF-1 α),vascular endothelial growth factor-A (VEGF-A) and vascular endothelial growth factor-D (VEGF-D)in hypoxic environment as well as the relationship between HIF-lα and VEGF-D.Methods Human esophageal cancer cell line EC9706 was cultured under hypoxia environment for 6,12 and 24 h,the cell radiosensitivity was evaluated by survival curve.HIF-1 α siRNA was constructed and transfected into human EC9706 cells.Protein expressions of HIF-1 α,VEGF-A and VEGF-D were analyzed by Western blot before and after RNA interference.Results EC9706 cells under hypoxia showed radioresistance with a SF2 of 0.62 higher than that of normoxic cells of 0.43.Moreover,the protein expressions of HIF-1α,VEGF-A and VEGF-D were all increased (F =205.24,227.88,130.55,P ＜0.05) due to hypoxia treatment.On the contrary,after HIF-1α siRNA transfer,the protein expressions of HIF-1α,VEGF-A and VEGF-D in EC9706 cells were not influenced by hypoxia treatment.Conclusions EC9706 cells in hypoxic environment was radioresistance,and the upexpressions of HIF1α,VEGF-A and VEGF-D may be involved.

Relationships between Lymph Node Metastasis and Expression of CD31, D2-40, and Vascular Endothelial Growth Factors A and C in Papillary Thyroid Cancer

OBJECTIVES: To investigate the relationships between lymph node metastasis (LNM) and expression of CD31, D2-40 and vascular endothelial growth factors (VEGF)-A and -C in patients with papillary thyroid cancer (PTC). METHODS: Paraffin-embedded thyroid tissues of 72 patients were evaluated, which included 25 patients with thyroid nodular hyperplasia (TNH), 24 PTC patients without LNM, and 23 PTC patients with LNM. Three pathologists, who were blinded to the patient's clinical information, assessed the immunohistochemical staining results. The amount of expression was scored as high (>25% of cells stained) or low (0-25%). RESULTS: A higher level of VEGF-A expression was observed in the PTC groups regardless of LNM when compared to the group with TNH (91.3%, 79.2%, 4.0%, respectively). VEGF-C expression in the PTC with LNM group was significantly higher than the other two groups (P<0.05). No difference in microvessel density (MVD) scores was observed using CD31 among the three groups. The lymphatic vessel density (LVD) score using D2-40 was significantly higher in patients having PTC with LNM than the other groups (P<0.05). CONCLUSION: VEGF-C and D2-40 were more highly expressed in patients having PTC with LNM than in patients having PTC without LNM or in those having TNH. Analysis of VEGF-C level and LVD using D2-40 may be helpful in the diagnosis of PTC and the evaluation of LNM potential in patients with PTC.