Objective: To determine the antiangiogenic activity of Telescopium telescopium (Horn snail) extract using in ovo chorioallantoic membrane (CAM) assay. Methods Methanolic extract of Telescopium telescopium was subjected to modified Kupchan partitioning. Four treatment groups – negative control, positive control (quercetin), test samples, and blanks – were used for the in ovo chorioallantoic membrane (CAM) assay. ImageJ software was used to measure average vessel diameter (DV) and total length (LT) to determine the degree of vascularization, percent inhibition, and antiangiogenic activity. Biochemical screening was done for the crude extract and the fraction with the highest percent inhibition.
Chorioallantoic Membrane ; Gastropoda

OBJECTIVETo explore the invasiveness of xenografts on chicken embryo chorioallantoic membrane (CAM) after tissue inhibitor of metalloproteinase-2 (TIMP-2) gene transfection.

METHODSFresh ameloblastoma tissues were minced into 1-2 mm3 and transplanted on the CAM. There were three groups named as control group (Empt), plasma transfection group (Lipo), and TIMP-2 gene transfection group (P). The specimens were respectively investigated by microscope indifferent spots after implanting. The volume of the xenografts and the weight of xenografts in the termination time of the experiment were recorded. The invasiveness of xenografts was divided into four grades by pathological examination. Western blot analysis was performed to investigate matrix metalloproteinase-2 (MIMP-2) and TIMP-2 protein in xenografts.

RESULTSAmeloblastoma tissues can survive on CAM and the tumor cells may invade it on 5-7 days after implanting. At 9 d after implanting, the invasiveness grades in P group were 7 in grade 0, 1 in grade 2, 0 in grade 3. The expression of TIMP-2 protein in P group was significantly higher than that in Empt group (P < 0.05). The expression of MMP-2 protein in P group was lower than that in Empt group (P < 0.05).

CONCLUSIONThe xenotransplanted tumor model of human ameloblastoma on CAM was successfully established. The invasiveness of ameloblastoma xenografts was suppressed might be due to TIMP-2 gene transfection.

Ameloblastoma ; Animals ; Chickens ; Chorioallantoic Membrane ; Heterografts ; Humans ; Matrix Metalloproteinase 2 ; Tissue Inhibitor of Metalloproteinase-2 ; Transfection

The MBT-2 mice bladder cancer tissues and the human bladder cancer tissues were implanted on the chorioallantoic membrance(CAM) of the immune deficient fertilized chicken eggs and the histopathologic changes of the CAM and gross morphologic changes of the implanted cancer tissues on CAM ere studied. The chemosensitivity tests using chicken CAM were performed for the 4 human bladder cancer tissues to mitomycin C, thiotepa and adriamycin. With this study, the following results were obtained: 1. The observation of the blood vessel on the chorioallantoic membrane was possible from the post-incubation 6th day group, but for the implantation of the cancer tissues, the blood vessels from the post-incubation 8th day group was appropriate. 2. The budding oif the host capillary vessel to the implanted cancer tissue were observed from the post-implantation second day. 3. The size of the post-implantation 7th day cancer tissues were varied from 2.3 to 9.2 folds to the size of the implantation day. 4. The total failure rate in experiment within post-operative 3rd day were 71.3 percent and the total failure rates in group who had the damage on the chorioallantoic membrance during operation was 82.5 percent. The failure rate of the experiment was declined acutely after post-operative 4th day. 5. The salvage of the eggs could be maintained until post-operative 7th day in 28.1 percent among chemosensitivity test group. 6. The 4 bladder cencer tissues which had the chemosensitivity test showed 1.6 to 7.1 fold growth to the inital implanted size and this meant resistance to the test drugs and these results were corresponded with clinical course.
Animals ; Blood Vessels ; Capillaries ; Chickens* ; Chorioallantoic Membrane* ; Doxorubicin ; Eggs* ; Humans* ; Mice* ; Mitomycin ; Ovum* ; Thiotepa ; Urinary Bladder Neoplasms* ; Urinary Bladder*

Eimeria (E.) tenella (local isolate) sporozoites were adapted on the chorioallantoic membrane (CAM) of 10-12 days chicken embryos and completed its life cycle in 6~7 days at 39 degrees C and 70 per cent humidity. Only 23 embryos (4.6%) were found dead from 1~4 day post inoculation of sporozoites with mild lesions on CAM with no gametocytes but few sporozoites in chorioallantoic fluid (CAF). On 5~7 day post inoculation, 432 embryos (86.4%) were found dead with severe haemorrhages on CAM and CAF contained uncountable number of gametocytes. After seven days post inoculation, 45 embryos (9%) were found to be alive. Some oocysts were also detected in the CAF on 6~7 days post inoculation. In the histological sections of the CAM, there were abundant small dark colored rounded bodies of gametes; distributed extensively in tissues of CAM on 5~7 days post inoculation of sporozoites. In some cases, cluster of small mature and immature relatively large bodies were seen in increasing numbers on 5~6 days post inoculation.
Animals ; Chick Embryo ; *Chickens ; Chorioallantoic Membrane/*parasitology ; Coccidiosis/parasitology/*veterinary ; Eimeria tenella/*growth&development ; Histocytochemistry ; Poultry Diseases/*parasitology

OBJECTIVETo investigate choline promoting angiogenesis on chick embryo chorioallantoic membrane (CAM).

METHODSCAM model was prepared, the choline chloride, human vascular endothelial growth factor (hVEGF) and normal saline were added respectively onto the carrier on the CAM, the state of angiogenesis was observed and the number of new blood vessels was counted.

RESULTSCholine chloride was tested at the concentrations of 0.5 nmol/L - 1 mmol/L in this experiment, when its concentrations were increased to 0.01 micromol/L - 1 000 micromol/L, it could stimulate angiogenesis, the minimum effective concentration was tested as 0.01 micromol/L, and its effect for promoting the angiogenesis was equivalent to that of hVEGF, the potent stimulator for angiogenesis.

CONCLUSIONThe result shows that choline can promote angiogenesis in the chick embryo chorioallantoic membrane.

Animals ; Chick Embryo ; blood supply ; drug effects ; Choline ; pharmacology ; Chorioallantoic Membrane ; blood supply ; drug effects ; Neovascularization, Physiologic ; drug effects

PURPOSE: Matrix metalloproteinases (MMPs) have been reported to play critical roles in the endothelial cell migration and matrix remodeling during angiogenic process. To investigate the roles of the membrane type MMP (MT1-MMP) by the matrix remodeling of endothelial cells, MT1-MMP expression vector was transfected into bovine aortic endothelial cells (BAECs). Increased ex+pression of MT1-MMP in BAECs enhanced the activation of MMP-2, invasion and migration of BAECs. Moreover, the capacity of tube formation was increased by MT1-MMP transfectants. These observations indicate that MT1-MMP is involved in the angiogenic process of endothelial cells in vitro. In this study, we attempted these effects were confirmed in vivo system. MATERIALS AND METHODS: In this study, we used MT1- MMP or Antisense MT1-MMP stable transfectants in HT1080 human fibrosarcoma cells. Chorioallantoic membrane (CAM) assay was used for the detection of angiogenesis in vivo and modified CAM assay for quantification of invasion of MT1-MMP transfected cells. RESULTS: In CAM assay, the formation of microvessels was stimulated by MT1-MMP transfectants. Invasive capacity of HT1080 cells was also increased in a novel in vivo metastasis model, PCR based CAM assay. CONCLUSION: These results identify the function of MT1- MMP during the neovascularization process.
Chorioallantoic Membrane* ; Endothelial Cells ; Fibrosarcoma ; Humans ; Matrix Metalloproteinase 1* ; Matrix Metalloproteinase 14 ; Matrix Metalloproteinases ; Membranes ; Microvessels ; Neoplasm Metastasis ; Polymerase Chain Reaction

It is generally accepted that a vascular accident in utero during the fetal period plays an important role in the occurrence of intestinal atresia. An experimental study, making use of chick embryos was made to induce intestinal atresia by vascular occlusion or ligation of a loop of intestine. A study using the chick embryos made it relatively easy to obtain the experimental group. Its advantages are a short incubation period(21 days) and many operations can be performed with only a limited supply of surgical instruments. Physiologic umbilical hernia of the chick embryo is present from the 9th to the 18th day of development. We used chick embryos between 9th to 15th day of development to perform experiments. In group I, as a control group, round shaped opening was made in the eggshell, shell membrane and chorioallantoic membrane with diameter of 1 cm, and then closed with transparent tape. In group II, the mesenteric artery was ligated with prolene 7-0. In the group III, a loop of intestine was ligated with prolene 7-0. The survival rate of group I was 35.7%(50/140), group II, 5.1%(36/700) and group III was 7.6%(53/700)(p<0.001). The intestinal atresia in hatched embryos showed no case in group I, 14cases out of 36cases in Group II(type II 5cases, type III 9cases), and all cases in Group III(type I 3cases, type II 29cases, type III 21cases). There was no significant relation between experimental group and type of intestinal atresia(p=0.09). In this experiments, the survival rate and incidence of intestinal atresia of group III were higher than group II. We concluded that vascular accident of intestine during fetal period was a factor in development of intestinal atresia, but, mechanical obstruction of intestinal loop was more important.
Animals ; Chick Embryo* ; Chorioallantoic Membrane ; Embryonic Structures ; Hernia, Umbilical ; Incidence ; Intestinal Atresia* ; Intestines ; Ligation ; Membranes ; Mesenteric Arteries ; Polypropylenes ; Surgical Instruments ; Survival Rate

Vascular endothelial growth factor-D (VEGF-D) and vascular endothelial growth factor receptor-2, -3 (VEGFR-2, -3) with their corresponding signaling pathway play significant roles in the development of the embryonic vascular system and pathological lymphangiogenesis. The study was aimed to express and purify the GST-VEGF-D fusion protein, and to explore the angiogenesis effect of VEGF-D. The total RNA was extracted from human fetal lung tissue, and the mature form of VEGF-D was expanded by polymerase chain reaction (PCR), then the plasmid pGEX-5X-1/VEGF-D was reconstructed and the GST-VEGF-D fusion protein expressed in transformed E.coli BL21-DE3. The results showed that the molecular mass of this fusion protein was 38 kD and compassed more than 15% of the total bacteria proteins. The fusion protein was recognized by anti-GST and anti-VEGF-D antibodies. The soluble GST-VEGF-D fusion protein could interact with VEGFR-3/Fc and was able to stimulate the proliferation of human erythroleukemia cell line (HEL) cells. The data of chick embryo chorioallantoic membrane (CAM) experiments indicated that GST-VEGF-D could induce the CAM angiogenesis. It is concluded that the GST-VEGF-D fusion protein with biological activity was successfully expressed, and which may provide an experimental model for the investigation of the VEGF-D-induced angiogenesis and lymphangiogenesis.
Animals ; Chick Embryo ; Chorioallantoic Membrane ; blood supply ; Humans ; Neovascularization, Physiologic ; drug effects ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology ; Vascular Endothelial Growth Factor D ; biosynthesis ; genetics ; pharmacology

Fuantai-03(FAT-03), isolated from the Dasyatis akajei, has a strong antiangiogenic activity. The recombinant Fuantai-03 (GST/rFAT-03) fusion protein can be obtained with the DNA recombination technology. In this study, expression conditions of GST/rFAT-03 were optimized by response surface experimental design method. The constructed engineering bacteria containing GST/rFAT-03 plasmid was induced by isopropy-beta-D-thiogalactosid (IPTG), the GST affinity column was used for isolation and purification, and then the effects of different culture time, IPTG concentration, induction temperature and induction time on the amount of soluble GST/rFAT-03 fusion protein were compared. The culture time for optimal expression was 6.13 h, IPTG concentration was 0.36 mmol/L, induction temperature was 19.71 degrees C, and induction time was 13.60 h. The amount of soluble GST/rFAT-03 fusion protein was 7.57 mg/L under above mentioned expression conditions. The results also showed that rFAT-03 significantly inhibited angiogenesis in chicken chorioallantoic membrane in a dose-dependent manner. Moreover, the soluble form of the target protein is useful for further work on purification and on studying its biological function.
Angiogenesis Inhibitors ; biosynthesis ; genetics ; Animals ; Chickens ; Chorioallantoic Membrane ; blood supply ; Escherichia coli ; genetics ; metabolism ; Fish Proteins ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Skates (Fish)

OBJECTIVETo investigate the effect of small interference RNA (siRNA) targeting nuclear factor-kappaB (NF-kappaB) on endometriosis.

METHODThe eutopic endometrium of women with endometriosis were transplanted into the nonvascular region of 8-day-old chicken embryo chorioallantocic membrane (CAM), and the effects of NF-kappaB p65 siRNA on the vascularization and endometriotic lesion formation were tested with proper controls.

RESULTSTransplantation of the endometrium onto the CAM resulted in a strong angiogenic response in the chicken tissue. The angiogenesis was significantly reduced and endometriotic lesion formation significantly suppressed with siRNA targeting NF-kappaB in comparison with the control group.

CONCLUSIONSThe NF-kappaB pathway is involved in the development of endometriotic lesions in vitro, and NF-kappaB gene silencing reduces endometriotic angiogenesis and promotes cell apoptosis in the endometriotic lesions, suggesting that NF-kappaB might be a good target for endometriosis treatment.

Animals ; Chick Embryo ; Chorioallantoic Membrane ; blood supply ; metabolism ; Endometriosis ; genetics ; physiopathology ; Female ; Humans ; NF-kappa B ; deficiency ; genetics ; Neovascularization, Pathologic ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics