1.Expressions of VEGF and Flt-1 in the testis, epididymis and epididymal sperm of adolescent rats.
Qing-yan AI ; Hong TIAN ; Jie ZHANG ; Li MA ; Nai-zhou MIAO ; Yong-wei HUO ; Li-rong WANG ; Shu-dong QIU ; Qiu-yang ZHANG
National Journal of Andrology 2008;14(10):871-875
OBJECTIVETo study the expressions of the vascular endothelial growth factor (VEGF) and its receptor fms-like tyrosine kinase-1 (Flt-1) in the testis, epididymis and epididymal sperm of adolescent rats and explore the functions of both the proteins in the male reproductive system.
METHODSThe expressions of VEGF and Flt-1 were detected in 20 adolescent SD rats, immunohistochemical staining used for both the testis and the epididymis and immunofluorescent staining for sperm.
RESULTSVEGF and Flt-1 proteins were specifically present in the testis, epididymis and sperm. In the testis, VEGF immunoreactive particles were localized in the cytoplasm of spermatogenic cells, the developing acrosome of spermatids, Sertoli cells and Leydig cells, while Flt-1 expressed mainly in the developing acrosome of spermatids and Leydig cells. In the epididymis, the cell-specific and region-specific expressions of VEGF and Flt-1 proteins were observed in the principal cells of epididymal epithelia, VEGF in the whole epididymis, while Flt-1 only in the caput and cauda segments. Both VEGF and Flt-1 were localized in the acrosome of the sperm head as well as in the neck, middle and principal segments of the sperm tail.
CONCLUSIONThe specific expression patterns of VEGF and Flt-1 in the rat testis, epididymis and sperm indicate that they may independently or collectively affect spermatogenesis and spermiotelcosis in either an autocrinological or a
Animals ; Epididymis ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Sexual Maturation ; Spermatozoa ; metabolism ; Testis ; metabolism ; Vascular Endothelial Growth Factor A ; biosynthesis ; Vascular Endothelial Growth Factor Receptor-1 ; biosynthesis
2.Inhibition of leukemic cell proliferation by human soluble VEGF-R1.
Jing ZHU ; Yan-Qiu SONG ; Wei LI ; Guan-Jun WANG
Journal of Experimental Hematology 2007;15(1):168-174
The current study was purposed to investigate the inhibitory effect of human soluble vascular endothelial growth factor-1 (sFLT-1) on the proliferation of leukemic cells in vitro. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the VEGF mRNA and VEGF-R1 (FLT-1) mRNA in K562, HL60, U937 leukemic cell lines and bone marrow LTC-IC. Flow cytometry was used to detect the VEGF and VEGF-R1 (FLT-1) in all above-mentioned cells. VEGF concentrations in the cell culture supernatants were determined by enzyme-linked immunosorbent assay (ELISA). Cell proliferation was determined by MTT after adding sFLT-1 to K562, HL60 and LTC-IC culture system. The result showed that expression of VEGF could be detected in K562, HL60, U937 leukemic cell lines and LTC-IC, especially K562, K562 and HL60 cell lines also expressed FLT-1, but a little expression was found in U937 and LTC-IC. sFLT-1 could effectively inhibit the growth of K562 and HL60 cell lines in dose-dependent manner. The highest inhibition rate was found at 48 hours after adding sFLT-1. It is concluded that sFLT-1 can inhibit the growth of some leukemic cell lines, and the inhibition effect enhances as the concentration of the sFLT-1 increase, but sFLT -1 not influence the proliferation of normal marrow cells.
Cell Proliferation
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drug effects
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HL-60 Cells
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Humans
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K562 Cells
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RNA, Messenger
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biosynthesis
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genetics
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U937 Cells
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Vascular Endothelial Growth Factor A
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antagonists & inhibitors
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biosynthesis
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genetics
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Vascular Endothelial Growth Factor Receptor-1
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biosynthesis
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genetics
;
physiology
3.Effect of rhodiola on expressions of Flt-1, KDR and Tie-2 in rats with ischemic myocardium.
Jian LI ; Wei-hu FAN ; Hong AO
Chinese Journal of Integrated Traditional and Western Medicine 2005;25(5):445-448
OBJECTIVETo investigate the effect of rhodiola on expression of vascular endothelial growth factors receptors (VEGFR) in myocardium of rats after myocardial infarction.
METHODSOn the basis of successful establishment of myocardial infarction rat model, the experimental animals were divided into the model group, the rhodiola group, the positive control group and the sham-operated group, they were sacrificed after 6 weeks feeding. Their hearts were resected and embedded in paraffin to make sections with standard immunohistochemistry stain. Then the stained slices were analyzed in the IMS cell imagine analysis system using immunohistochemical quantitative analysis software. The field of vision of left ventricular myocardial tissue in three sites selected from the marginal area of infarction in each slice were determined, the mean value was then converted to positive area. Meanwhile, the mean optical density (OD) was calculated and the various expressions of VEGFR, i.e. Flt-1, KDR and angiopoietin receptor (Tie-2) were measured.
RESULTSThe expressions of Flt-1 and Tie-2 in myocardial tissue were significantly increased in the rhodiola treated group after treatment, showing significant difference as compared with those in the positive control group and the model group (P < 0.05). The expression of KDR in myocardium after rhodiola intervention was higher than that in the sham-operated and nonintervened group (P < 0.05), but insignificantly different to that in the positive control group and model group.
CONCLUSIONRhodiola could improve angiogenesis to ameliorate myocardial ischemia by regulating the expression of Flt-1 and Tie-2 in ischemic myocardium.
Animals ; Drugs, Chinese Herbal ; pharmacology ; Male ; Myocardial Infarction ; metabolism ; Myocardium ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptor, TIE-2 ; biosynthesis ; genetics ; Receptors, Vascular Endothelial Growth Factor ; biosynthesis ; genetics ; Rhodiola ; Vascular Endothelial Growth Factor Receptor-1 ; biosynthesis ; genetics ; Vascular Endothelial Growth Factor Receptor-2 ; biosynthesis ; genetics
4.Construction and expression of soluble vascular endothelial growth factor receptor-1 eukaryotic expression vector and its effect on proliferation of vascular endothelial cells.
Lingling JI ; Shuhua MAO ; Hong LIU ; Shiye XU ; Yu YANG ; Cheng YI ; Ying HUANG
Journal of Biomedical Engineering 2010;27(2):369-372
This study sought to construct recombinant eukaryotic plasmid pcDNA3. 1-sFlt-1 and observe its effect on proliferation of vascular endothelial cells. Total RNA was extracted from human umbilical vein endothelial cells (HUVECs) firstly. The 1st-3rd Ig-like domains of Flt were amplified by polymerase chain reaction (PCR) from the full-length cDNA. Subsequently, the PCR product was cloned into the eukaryotic plasmid pcDNA3.l(+)/myc-His. The constructed recombinant plasmid pcDNA3. 1-sFlt-1 was sequenced. Then recombinant plasmid was transfected into Lewis lung cancer cells. RT-PCR and SDS-PAGE were used to detect the expression of soluble vascular endothelial growth factor (VEGF) receptor mRNA and protein, respectively. MTT method was used to evaluate the effect of sFlt-1 protein on proliferation of HUVECs induced by VEGF. The results showed: (1) The sequence of inserted fragment was correct. (2) Lewis lung cancer cells with recombinant plasmid transfection could express the soluble VEGF receptor mRNA and protein stably. (3) Culture supernatant of Lewis lung cancer cells with sFlt-1 could significantly inhibit the proliferation of HUVECs induced by VEGF. These data suggested that recombinant eukaryotic plasmids pcDNA3. 1-sFlt-1 was constructed successfully, sFlt-1 mRNA and protein were expressed in eukaryotic system stably and sFlt-1 protein could significantly inhibit the proliferaton of endothelial cells induced by VEGF.
Cell Line, Tumor
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Cell Proliferation
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drug effects
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Cells, Cultured
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Genetic Vectors
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genetics
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Human Umbilical Vein Endothelial Cells
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cytology
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Humans
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Transfection
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Vascular Endothelial Growth Factor A
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pharmacology
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Vascular Endothelial Growth Factor Receptor-1
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biosynthesis
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genetics
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pharmacology
5.Inhibition of expression of vascular endothelial growth factor and its receptors in pulmonary adenocarcinoma cell by TNP-470 in combination with gemcitabine.
Xue-fen WANG ; Ling-fang TU ; Li-hong WANG ; Jian-ying ZHOU
Journal of Zhejiang University. Science. B 2006;7(10):837-843
Angiogenesis is required for solid tumor growth and facilitates tumor progression and metastasis. The inhibition effects of O-(chloroacetyl-carbamoyl) fumagillol (TNP-470), an angiogenesis inhibitor, and gemcitabine, a chemotherapeutic agent, on expression of growth factors were investigated using human pulmonary adenocarcinoma cell line, A549. The A549 cells were divided into four groups: control group, 10(-6) mg/ml gemcitabine treated group, 10(-4) mg/ml TNP-470 treated group and gemcitabine+TNP-470 treated group. The mRNA and protein expression of vascular endothelial growth factor (VEGF) and its receptors, FMS-like tyrosine kinase-1 (FLT-1) and kinase insert domain-containing receptor (KDR), in different groups were measured. The growth of A549 cell cultured with gemcitabine or TNP-470 was inhibited in an almost dose-dependent manner. Although gemcitabine (10(-6) mg/ml) alone and TNP-470 (10(-4) mg/ml) alone had no effect on the mRNA and protein expression of VEGF and its receptors (FLT-1, KDR) in A549 cells compared to the control (P>0.05), 10(-6) mg/ml gemcitabine in combination with 10(-4) mg/ml TNP-470 had significant effect (P<0.01). Moreover, combination of the two drugs significantly inhibited the mRNA expression of VEGF, FLT-1 and KDR compared to either drug alone (P<0.05). This study suggests that combined treatment with TNP-470 plus gemcitabine may augment the antiangiogenic and antineoplastic effects in lung cancer cells in vitro.
Adenocarcinoma
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drug therapy
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Antineoplastic Combined Chemotherapy Protocols
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therapeutic use
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Cell Line, Tumor
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Cell Proliferation
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Cyclohexanes
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administration & dosage
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Deoxycytidine
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administration & dosage
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analogs & derivatives
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Disease Progression
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Gene Expression Regulation, Neoplastic
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Humans
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Lung Neoplasms
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drug therapy
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Neoplasm Metastasis
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Protein Structure, Tertiary
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Sesquiterpenes
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administration & dosage
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Vascular Endothelial Growth Factor A
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biosynthesis
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Vascular Endothelial Growth Factor Receptor-1
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biosynthesis
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Vascular Endothelial Growth Factor Receptor-2
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biosynthesis
6.Expression of vascular endothelial growth factor and its receptors KDR and Flt1 in acute myeloid leukemia.
Yi WANG ; Zhi-jian XIAO ; Peng LIU ; Chen YANG ; Ren-chi YANG ; Ying-lin CAI ; Zhong-chao HAN
Chinese Journal of Hematology 2003;24(5):249-252
OBJECTIVETo evaluate the expression of vascular endothelial growth factor (VEGF) and its receptors KDR and Flt1 in patients with acute myeloid leukemia (AML).
METHODSThe expression of VEGF and its receptors mRNA was assayed by RT-PCR, the plasma of VEGF by ELISA.
RESULTSIn 13 AML cell lines, the expression of VEGF, KDR and Flt1 mRNA were found in 13 (100%), 7 (53.8%) and 12 (92.3%), respectively. There were 21 (65.6%), 1 (3.1%), and 17 (53.1%) of 31 (AML) patients bone marrow mononuclear cells (BMMNCs) expressing VEGF, KDR and Flt1 mRNA, respectively. None of BMMNCs from 3 normal donor and CD(34)(+) cells from 2 normal donor was found to express VEGF, KDR and Flt1 mRNA. The plasma level of VEGF of 39 patients (new diagnosed, relapsed and secondary-AML) before treatment was (135.3 +/- 87.9) ng/L which was significantly higher than that of 15 complete remission (CR) patients (80.6 +/- 36.9) ng/L and 12 normal donors (80.6 +/- 33.1) micro g/L (P = 0.028, 0.007). The plasma level of VEGF of 15 non-responsive patients was (188.2 +/- 118.6) ng/L after two cycles of chemotherapy which was higher than that of 20 CR patients [(104.2 +/- 30.9) ng/L] (P = 0.004).
CONCLUSIONVEGF and its receptors KDR and Flt1 mRNAs were expressed in BMMNCs of AML patients. The plasma level of VEGF directly affected the response to chemotherapy in AML patients.
Enzyme-Linked Immunosorbent Assay ; Humans ; Leukemia, Myeloid, Acute ; drug therapy ; metabolism ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics ; Vascular Endothelial Growth Factor Receptor-1 ; biosynthesis ; genetics ; Vascular Endothelial Growth Factor Receptor-2 ; biosynthesis ; genetics
7.Expression of VEGF in prostate cancer and its correlation with ET-1.
Wen-Quan ZHOU ; Hong-Lin YIN ; Zheng-Yu ZHANG ; Xiao-Ming YI ; Jing-Ping GE ; Shui-Gen ZHOU ; Wen CHENG ; Wu WEI ; Hong-Qing MA ; Heng-Hui MA ; Jian-Ping GAO
National Journal of Andrology 2008;14(11):987-992
OBJECTIVETo investigate the expressions of VEGF in prostate cancer (PCa) and benign prostatic hyperplasia (BPH), their clinical significance and their relationship with that of ET-1.
METHODSA total of 44 specimens of PCa and 36 of BPH tissues were examined by the immunohistochemical Elivision plus method for the expressions of VEGF and ET-1. The intensity of staining for VEGF and ET-1 was assessed by light microscopy on a scale from "-" to "+ + +".
RESULTSThe rates of positive expression of VEGF were 69.4% in BPH and 80.9% in PCa, positive staining mostly in the cytoplasm of glandular epithelia and cancer cells, and strongly positive in all the stroma vascular endothelial cells. The staining intensity of VEGF was significantly higher in the PCa than in the BPH group (P < 0.05) , in the bone metastasis (BM) than in the non-BM group (P < 0.01), and in the lowly than in the highly and moderately differentiated PCa tissues (P < 0.01). The expression of VEGF was positively correlated with that of ET-1 ( r(s) = 0.780, P < 0.01).
CONCLUSIONVEGF is involved in the development, progression and metastasis of PCa. VEGF and ET-1 may play a joint role in its development and progression.
Aged ; Aged, 80 and over ; Endothelin-1 ; biosynthesis ; Humans ; Male ; Middle Aged ; Neoplasm Metastasis ; Neoplasm Staging ; Prostatic Hyperplasia ; metabolism ; pathology ; Prostatic Neoplasms ; metabolism ; pathology ; Vascular Endothelial Growth Factor Receptor-1 ; biosynthesis
8.Effect of phosgene on apoptosis of alveolar type II cells and vascular endothelial growth factor in exposed mice.
Wen-li LI ; Chun-xu HAI ; Xu-jun QIN ; Xin LIANG ; Hong-li CHEN
Chinese Journal of Industrial Hygiene and Occupational Diseases 2004;22(3):197-199
OBJECTIVETo study the apoptosis of alveolar type II cells, alterations of vascular endothelial growth factor (VEGF), VEGF receptor (Flt1) in serum and lung and expression of VEGF mRNA in lung in pulmonary edema mice induced by phosgene.
METHODSTwenty-six BALB/C mice were randomly divided into 2 groups: control group, exposed group (13 mice in each group). Mice of exposed group were intoxicated by inhalation of phosgene 11.9 mg/L for 5 minutes. Mice of control group were treated as the same way by inhalation of air. Isolation of mice alveolus type II cells 4 h after intoxication was carried out to observe their apoptosis under electron microscope. Contents of VEGF and Flt1 in lung and serum by ELISA, and expression of VEGF mRNA were determined.
RESULTSAlveolar type II cells were identified by tannic acid staining and electron microscopy. After exposed to 11.9 mg/L of phosgene for 5 minutes, the apoptotic body in alveolus type II cells was found in exposed group. The contents of VEGF in serum and lung and Flt1 in lung of exposed mice [(134.07 +/- 120.26), (477.76 +/- 98.06), (1,2818.48 +/- 2,304.15) pg/ml] were significantly lower than those of control group [(445.57 +/- 173.30), (1,026.87 +/- 474.56), (21,976.51 +/- 7,421.01) pg/ml, P < 0.05] but the content of Flt1 in serum [(2,369.56 +/- 381.70) pg/ml] was higher than that in control group [(1,898.00 +/- 453.69) pg/ml, P < 0.05]. The expression of VEGF mRNA in pulmonary edema mice was decreased.
CONCLUSIONPhosgene can induce apoptosis of alveolar type II cells, and decrease in the content of VEGF and Flt1, and expression of VEGF mRNA in lung.
Animals ; Apoptosis ; drug effects ; Cells, Cultured ; Chemical Warfare Agents ; toxicity ; Endothelial Growth Factors ; biosynthesis ; genetics ; Enzyme-Linked Immunosorbent Assay ; Male ; Mice ; Mice, Inbred BALB C ; Phosgene ; toxicity ; Pulmonary Alveoli ; drug effects ; metabolism ; pathology ; Pulmonary Edema ; chemically induced ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Vascular Endothelial Growth Factor A ; analysis ; genetics ; physiology ; Vascular Endothelial Growth Factor Receptor-1 ; analysis ; genetics
9.Increased sFlt-1 to PlGF Ratio in Women Who Subsequently Develop Preeclampsia.
Shin Young KIM ; Hyun Mee RYU ; Jae Hyug YANG ; Moon Young KIM ; Jung Yeol HAN ; Joo Oh KIM ; Jin Hoon CHUNG ; So Yeon PARK ; Moon Hee LEE ; Do Jin KIM
Journal of Korean Medical Science 2007;22(5):873-877
The purpose of this study was to determine whether the levels of soluble fms-like tyrosine kinase-1 (sFlt-1) and placenta growth factor (PlGF) are altered during the second trimester in the plasma of women who subsequently develop preeclampsia. We performed a case-control study to compare the levels of sFlt-1 and PlGF in the preeclamptic (n=46) and normal pregnant women (n=100). The maternal plasma levels of sFlt-1 and PlGF were measured by enzyme-linked immunosorbent assay. The sFlt-1 levels were significantly higher in the preeclamptic women than in normal controls (p<0.001), while the PlGF levels were significantly lower (p<0.001). In normal controls, sFlt-1 levels were positively correlated (r=0.27, p=0.008), whereas, in the preeclamptic women, those were negatively correlated with the PlGF levels (r=-0.423, p=0.005). Furthermore, the log[sFlt-1/PlGF] ratio was significantly higher in the preeclamptic women than in normal controls (p<0.001). The receiver operating characteristic curve revealed a specificity of 78% with a diagnostic sensitivity of 80.4%; the optimal cut-off value of the log[sFlt-1/PlGF] ratio was 1.4 (95% CI 0.756-0.910, p<0.001). Preeclampsia showed a strong association with increased levels of sFlt-1 and decreased levels of PlGF in the second trimester maternal plasma. Accordingly, the sFlt-1/PlGF ratio may provide early prediction of subsequent development of preeclampsia.
Adult
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Biological Markers/metabolism
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Case-Control Studies
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Female
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Humans
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Immunoassay
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Middle Aged
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Placenta/metabolism
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Pre-Eclampsia/*diagnosis/*metabolism
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Pregnancy
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Pregnancy Proteins/*biosynthesis
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Pregnancy Trimester, Second
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ROC Curve
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Sensitivity and Specificity
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Vascular Endothelial Growth Factor Receptor-1/*biosynthesis
10.Effect of Endogenous Bone Marrow Derived Stem Cells Induced by AMD-3100 on Expanded Ischemic Flap.
Hii Sun JEONG ; Hye Kyung LEE ; Kwan Chul TARK ; Dae Hyun LEW ; Yoon Woo KOH ; Chul Hoon KIM ; In Suck SEO
Journal of Korean Medical Science 2014;29(Suppl 3):S237-S248
The purpose of this study was to devise an expanded ischemic flap model and to investigate the role of AMD-3100 (Plerixafor, chemokine receptor 4 inhibitor) in this model by confirming its effect on mobilization of stem cells from the bone marrow. Male Sprague-Dawley rats were used as an animal research model. The mobilization of stem cells from the bone marrow was confirmed in the AMD-3100-treated group. The fractions of endothelial progenitor cells (EPC) and the vascular endothelial growth factor receptor (VEGFR) 2+ cells in the peripheral blood were increased in groups treated with AMD-3100. The expression of vascular endothelial growth factor (VEGF) was increased in response to expansion or AMD injection. The expression of stromal cell derived factor (SDF)-1 and VEGFR2 were increased only in unexpanded flap treated with AMD-3100. Treatment with AMD-3100 increased both the number and area of blood vessels. However, there were no statistically significant differences in the survival area or physiologic microcirculation in rats from the other groups. This endogenous neovascularization induced by AMD-3100 may be a result of the increase in both the area and number of vessels, as well as paracrine augmentation of the expression of VEGF and EPCs. However, the presence of a tissue expander under the flap could block the neovascularization between the flap and the recipient regardless of AMD-3100 treatment and expansion.
Animals
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Anti-HIV Agents/pharmacology
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Bone Marrow Cells/cytology
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Chemokine CXCL12/biosynthesis
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Endothelial Progenitor Cells/*cytology
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Hematopoietic Stem Cells/*cytology
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Heterocyclic Compounds/*pharmacology
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Hypoxia-Inducible Factor 1, alpha Subunit/metabolism
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Male
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Neovascularization, Physiologic
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Nitric Oxide Synthase Type III/metabolism
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Rats
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Rats, Sprague-Dawley
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Receptors, CXCR4/antagonists & inhibitors
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Surgical Flaps/*blood supply/surgery
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Tissue Expansion/*methods
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Vascular Endothelial Growth Factor A/biosynthesis
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Vascular Endothelial Growth Factor Receptor-2/biosynthesis/metabolism