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OBJECTIVE@#To compare the clinical effect of lower extremity varicose veins between fire needling bloodletting and operation, and to explore the possible mechanism.@*METHODS@#A total of 60 patients were randomized into an observation group and a control group, 30 cases in each one. In the control group，the operation was adopted. The fire needling bloodletting was applied in the observation group, twice a week for 4 weeks. Before and after treatment, the venous clinical severity score (VCSS) and venous disability score (VDS) were recorded, the hemorheological indexes [blood viscosity, plasma viscosity, hematocrit, fibrinogen and erythrocyte sedimentation rate (ESR)], immune inflammatory response indexes[serum C-reactive protein (CRP), tumor necrosis factor-α(TNF-α) and interleukin-6 (IL-6)], vascular endothelial cell function indexes [the number of circulatingendothelial cell (CEC), plasma endothelin (ET-1) and NO)] and apoptosis indexes (Bcl-2, Bax and Caspase-3) were detected in the two groups.@*RESULTS@#Compared before treatment, the scores of VCSS and VDS, hemorheological indexes, immune inflammatory response indexes and levels of plasma NO after treatment were reduced in the two groups (<0.05). The level of serum Bax after treatment was reduced in the observation group (<0.05). The number of CEC and levels of plasma ET-1 after treatment were increased in the two groups (<0.05). The levels of serum Bcl-2 and Caspase-3 after treatment were increased in the observation group (<0.05). In the observation group, the scores of VCSS and VDS, hemorheological indexes，immune inflammatory response indexes, vascular endothelial cell function indexes and level of serum Bax after treatment were lower than the control group (<0.05), and the levels of Bcl-2 and Caspase-3 were higher than the control group (<0.05).@*CONCLUSION@#Fire needling bloodletting could effectively treat lower extremity varicose veins, and the mechanism may be related to the improvement of hemorheology, downregulation of immune inflammatory response, improvement of vascular endothelial cell function and inhibition of apoptosis.
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OBJECTIVE@#To explore application of targeted contrast enhanced ultrasonography in diagnosis of early stage vascular endothelial injury and diabetic nephropathy.@*METHODS@#Targeted SonoVue-TM microbubble was prepared by attaching anti-TM monoclonal antibody to the surface of ordinary microbubble SonoVue by biotin - avidin bridge method and ultrasonic instrument was used to evaluate the developing situation of targeted microbubble in vitro. Twenty 12-week-old male GK rats and 20 Wistar rats were enrolled in this study, and were randomly divided into targeted angiography group and ordinary angiography group. Targeted microbubbles SonoVue-TM or general microbubble SonoVue were rapidly injected to the rats via tail vein; the developing situation of the two contrast agents in rats kidneys was dynamically observed. Time-intensity curve was used to analyze rat kidney perfusion characteristics in different groups.@*RESULTS@#Targeted ultrasound microbubble SonoVue-TM was successfully constructed, and it could be used to develop an external image. Targeted microbubbles SonoVue-TM enabled clear development of experimental rat kidney. Time-intensity curve shapes of rat kidney of the two groups showed as single apex with steep ascending and slowly descending branch. Compared with the control group, the rising slope of the GK rat renal cortex, medulla in targeted angiography group increased (P < 0.05); the peak intensity of medulla increased (P < 0.05), and the total area under the curve of medulla increased (P < 0.05). Compared with control group, the ascending branch of the GK rat in renal cortex, medulla in ordinary angiography group increased (P < 0.05). The peak intensity of the curve increased (P < 0.05), and the total area under the curve increased (P < 0.05). Compared with the ordinary angiography group, the peak of GK rat medulla curve in targeted angiography group intensity increased (P < 0.05), and the total area under the curve increased (P < 0.05).@*CONCLUSIONS@#Targeted microbubbles SonoVue-TM can make a clear development of experimental rat kidney, its stable performance meet the requirement of ultrasonic observation time limit, and it can reflect early changes of blood perfusion in GK rat kindey.
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Objective: To explore application of targeted contrast enhanced ultrasonography in diagnosis of early stage vascular endothelial injury and diabetic nephropathy. Methods: Targeted SonoVue-TM microbubble was prepared by attaching anti-TM monoclonal antibody to the surface of ordinary microbubble SonoVue by biotin - avidin bridge method and ultrasonic instrument was used to evaluate the developing situation of targeted microbubble in vitro. Twenty 12-week-old male GK rats and 20 Wistar rats were enrolled in this study, and were randomly divided into targeted angiography group and ordinary angiography group. Targeted microbubbles SonoVue-TM or general microbubble SonoVue were rapidly injected to the rats via tail vein; the developing situation of the two contrast agents in rats kidneys was dynamically observed. Time-intensity curve was used to analyze rat kidney perfusion characteristics in different groups. Results: Targeted ultrasound microbubble SonoVue-TM was successfully constructed, and it could be used to develop an external image. Targeted microbubbles SonoVue-TM enabled clear development of experimental rat kidney. Time-intensity curve shapes of rat kidney of the two groups showed as single apex with steep ascending and slowly descending branch. Compared with the control group, the rising slope of the GK rat renal cortex, medulla in targeted angiography group increased (P < 0.05); the peak intensity of medulla increased (P < 0.05), and the total area under the curve of medulla increased (P < 0.05). Compared with control group, the ascending branch of the GK rat in renal cortex, medulla in ordinary angiography group increased (P < 0.05). The peak intensity of the curve increased (P < 0.05), and the total area under the curve increased (P < 0.05). Compared with the ordinary angiography group, the peak of GK rat medulla curve in targeted angiography group intensity increased (P < 0.05), and the total area under the curve increased (P < 0.05). Conclusions: Targeted microbubbles SonoVue-TM can make a clear development of experimental rat kidney, its stable performance meet the requirement of ultrasonic observation time limit, and it can reflect early changes of blood perfusion in GK rat kindey.
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Background Choroidal neovascularization (CNV) is a main cause of visual impairment in many retinal diseases.To create an ideal CNV animal model is very important for the experimental and clinical study of CNV.The assessment method of repeatable and reliable for CNV model is still seldom.Objective This experiment was to explore the label value of fluorescent antibody for visualizing and quantifying the morphologic changes associated with laser-induced CNV.Methods Laser-induced CNV models were created in 30 eyes of 15 male SPF C57BL/6J mice by Krypton red laser irradiating fundus 2 spots around the optical disc with the wavelength 647.1nm,power 260 mW,spot diameter 50μm and exposure time 0.05 seconds.The CNV was evaluated at 5 minutes,4,7,14 and 28 days after laser injury by using fundus photography and fundus fluorescein angiography (FFA),and the successful models were identified as the rupture of Bruch's membrane.The mice were then immediately sacrificed and the eyeballs were enucleated to prepare the choroidal flatmounts.The posterior eye cups were fluorescently labeled with markers of cell nuclei (DAPI,4',6'-diamino-2-phenylindole),endothelial cells (isolectin-B4),and filamentous actin (phalloidin).The CNV areas from specimens were measured by Image pro plus 6.0.Two eyes from one matched mouse without receiving photocoagulation were used as the controlls.This study followed the Standard of Association for Research in Vision and Ophthalmology.Results No any CNV was seen in photocoagulated eyes in 5 minutes and 4 days after laser irradiation.The first sign of CNV appeared at 7 days following photocoagulation.The incidence of fluorescein leakage was 76.47% (26/34),81.81% (18/22),50.00% (5/10) at 7,14 and 28 days,respectively.The fluomicroscope examination showed that in unphotocoagulated areas,retinal pigment epithelial (RPE) cells were visualized with a uniform hexagonal array.Immediately after laser exposure,a circular area devoid of fluorescent labeling was observed,indicating disruption of the choroid-Bruch membrane-RPE complex.On the fourth day,cellular debris and fragmented nuclei were presented and an autofluorescent ring was visible at the site of Bruch's membrane disruption.The number of CNV vessels increased exponentially during the next 3 days.At 7 days,a well-defined isolectin-B4 labeled CNV network was exhibited and lasted for 28 days.The CNV areas were (7.99±0.42)×103μm2,(16.89±3.77)×103μm2,(14.37±4.02)×103μm2 at 7,14 and 48 days after photocoagulation respectively,showing a significant difference among these three groups (F=17.340,P=0.000),and the CNV area was significantly increased in the photocoagulating eyes in 14 days and 28 days compared with 7 days (q=16.46,q=15.54,P＜0.01).Conclusion Fluorescent antibody labeling allows the well identification and measurement of laser-induced CNV lesions in mouse choroid/RPE flatmounts.This technique offers excellent morphologic detail and facilitates the study of critical early events in CNV.CNV complexes are labeled at an early stage,providing a more accurate preclinical evaluation of antiangiogenic molecule.
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<p><b>BACKGROUND</b>Ginsenosides are main components extracted from ginseng, and ginsenoside Rg3 is one of the most important parts. Ginsenoside Rg3 has been found to inhibit several kinds of tumor growth and metastasis. The present study was undertaken to investigate the effect of ginsenoside Rg3 on human ovarian cancer metastasis and the possible mechanism.</p><p><b>METHODS</b>The experimental lung metastasis models of ovarian cancer SKOV-3 and the assay of tumor-induced angiogenesis were used to observe the inhibitory effects of Rg3 on tumor metastasis and angiogenesis. The effect of Rg3 on invasive ability of SKOV-3 cells in vitro was detected by Boyden chamber, and immunofluorescence staining was used to recognize the expression of matrix metalloproteinase 9 (MMP-9) in SKOV-3 cells.</p><p><b>RESULTS</b>In the experimental lung metastasis models of ovarian cancer, the number of tumor colonies in the lung and vessels oriented toward the tumor mass in each ginsenoside Rg3 group, was lower than that of control group. The invasive ability and MMP-9 expression of SKOV-3 cells decreased significantly after treatment with ginsenoside Rg3.</p><p><b>CONCLUSIONS</b>Ginsenoside Rg3 can significantly inhibit the metastasis of ovarian cancer. The inhibitory effect is partially due to inhibition of tumor-induced angiogenesis and decrease of invasive ability and MMP-9 expression of SKOV-3 cells.</p>
Subject(s)
Animals , Female , Humans , Mice , Cell Line, Tumor , Ginsenosides , Pharmacology , Lung Neoplasms , Matrix Metalloproteinase 9 , Metabolism , Neoplasm Invasiveness , Neovascularization, Pathologic , Ovarian Neoplasms , Drug Therapy , PathologyABSTRACT
<p><b>BACKGROUND</b>Ginsenoside Rg3, the main component isolated from ginseng, inhibits some kinds of tumour growth and angiogenesis. The combination of low dose chemotherapy and antiangiogenesis inhibitors suppresses growth of experimental tumours more effectively than conventional therapy. The effect of this combination on ovarian cancer remains to be evaluated. Therefore, we investigated the synergism of ginsenoside Rg3 and cyclophosphamide (CTX) on growth and angiogenesis of human ovarian cancer.</p><p><b>METHODS</b>Twenty-eight female athymic mice were divided randomly into 4 groups of 7: ginsenoside Rg3, CTX, ginsenoside Rg3 and CTX combination and control, after being transplanted with ovarian cancer cells (SKOV-3). The mice were given intraperitoneal injection of ginsenoside Rg3 and CTX for the 10 days following inoculation of SKOV-3 cells. The life quality and number of living days of mice were recorded. The size of tumour, tumour inhibitive rate, life elongation rate, proliferating cell nuclear antigen labelling index (PCNALI), expression of vascular endothelial cell growth factor (VEGF) and microvessel density (MVD) of the tumour tissues were estimated.</p><p><b>RESULTS</b>Life quality of mice in ginsenoside Rg3 and combined treatment groups were better and number of living days longer than control. Average tumour weights of each treated group were less than control and there was no significant difference among the treated groups. PCNALI of treated groups was lower than control. The MVD value and VEGF expression in treated groups were significantly lower than control and the MVD values of ginsenoside Rg3 and combined treatment groups were lower than that of CTX group.</p><p><b>CONCLUSIONS</b>Ginsenoside Rg3 significantly inhibited growth and angiogenesis of ovarian cancer when used alone or combined with CTX. Ginsenoside Rg3 and CTX combination reinforced the antitumour effect each other and improved the living quality and survival time of mice with tumour.</p>
Subject(s)
Animals , Female , Humans , Mice , Antineoplastic Combined Chemotherapy Protocols , Pharmacology , Cell Line, Tumor , Cell Proliferation , Cyclophosphamide , Ginsenosides , Immunohistochemistry , Neovascularization, Pathologic , Drug Therapy , Pathology , Ovarian Neoplasms , Drug Therapy , Pathology , Proliferating Cell Nuclear Antigen , Vascular Endothelial Growth Factor AABSTRACT
NOD mice were treated intraperitoneally with tripterygium wilfordii ployglycosidium (TWP) for 4 weeks to observe the incidence of cyclophosphamide accelerated diabetes.The apoptosis of?-cells was detected by TUNEL,the expression of caspase-3 in islets of the NOD mice was detected by immunohistochemistry and the expression of caspase-8 mRNA in pancreas by RT-PCR.The results revealed that the incidence of diabetes in TWP group was lower than that in control group.The apoptosis index of?-cells was decreased in TWP group. The expression of caspase-3 in islets and the expression of caspase-8 mRNA in pancreas in TWP group were lower than those in control group.