血管内皮素生长因子及内皮抑素的表达水平与肺肿瘤分期、转移和治疗的关系

肺癌位居各类肿瘤的首位,成为危害人类健康和生命的头号杀手。大部分肺癌一经临床确诊已处于中晚期,合并恶性胸腔积液和其他部位转移,治疗效果较差,预后不良。本文旨在探讨血管内皮素生长因子（vascular endothelial growth factor,VEGF）及内皮抑素（endostatin,ES）在血清中及癌性胸水中的表达水平,与非小细胞肺癌（non-small cell lung cancer,NSCLC）分期、转移和治疗的关系,了解相关的规律,为肿瘤诊断治疗提供可靠的依据。

刺芒柄花素对老年晚期肺癌患者胸水及血清中内皮抑素、VEGF、MMP-2、bFGF及肿瘤标志物的影响

目的探讨刺芒柄花素对老年晚期肺癌患者胸水及血清中内皮抑素(endostatin,ES)、血管内皮素生长因子(vascular endothelial growth factor,VEGF)、基质金属蛋白酶2(matrix metalloproteinases 2,MMP-2)、碱性成纤维细胞生长因子(basic fibroblast growth factor,b FGF)及肿瘤标志物的影响。方法选取河南省南阳市中心医院诊治的老年晚期肺癌患者67例,根据随机数字表法随机分为2组:对照组33例,行临床常规NP方案化疗;实验组34例,在对照组相同的临床常规治疗基础上加用刺芒柄花素予以治疗,比较2组患者胸水和血清中ES、VEGF、MMP-2、b FGF、肿瘤标志物水平以及生存质量评分变化。结果与对照组比较,实验组胸水和血清中ES、VEGF、MMP-2、b FGF含量均明显降低(P<0.05);实验组血清癌胚抗原(carcino embryonie antigen,CEA)、神经元特异性烯醇化酶(neuron specific enolase,NSE)、细胞角蛋白19片段(cytokeratin-19-fragments,CYFRA21-1)、糖类抗原125(carbohydrate antigen125,CA125)水平明显降低(P<0.05);实验组患者生存质量评分情况明显优于对照组(χ2=4.96,P<0.05)。结论刺芒柄花素对老年晚期肺癌患者治疗具有较好的临床疗效,且能有效改善患者生存质量,降低胸水和血清中ES、VEGF、MMP-2、b FGF等指标和肿瘤标志物水平,对临床治疗老年晚期肺癌具有重要意义。

FLT4调控羊驼黑色素细胞中黑色素生成

血管内皮素生长因子受体3(vascular endothelial growth factor receptor 3,VEGFR-3/FLT4)与黑色素瘤细胞的生长有关,其可能对毛色及黑色素生成有一定的调控作用。为探讨FLT4基因对羊驼毛色及黑色素生成的影响,本文采用免疫组织化学、qRT-PCR、Western印迹对FLT4在不同毛色羊驼皮肤毛囊中的表达进行了研究。结果显示,FLT4在不同毛色毛囊中外根鞘及毛乳头阳性表达不同,且棕色皮肤比白色皮肤阳性信号强;FLT4 mRNA在棕色羊驼皮肤的表达量明显高于白色羊驼皮肤(P<0.01);FLT4蛋白在棕色皮肤中的表达量明显高于白色皮肤(P<0.01);为进一步研究FLT4对黑色素生成的影响;通过对体外培养的羊驼皮肤黑色素细胞中添加不同浓度的FLT4蛋白(0,1,10,50,100 ng/m L),与对照组相比,添加FLT4蛋白后酪氨酸酶(TYR)、酪氨酸相关蛋白-1(TYRP1)、受体酪氨酸激酶c-KIT、小眼畸形相关转录因子(MITF)的mRNA和蛋白质表达明显增加,10 ng/m L组表达量最高(P<0.01);黑色素含量测定结果表明,不同浓度的FLT4蛋白处理的羊驼皮肤黑色素细胞的黑色素含量比对照组明显升高,添加FLT4蛋白10 ng/m L表达量最高(P<0.01)。由此可知,FLT4可以通过调节毛色相关基因的表达进而引起黑色素细胞中黑色素含量的变化。

Increased protein and mRNA expression of endostatin in the ischemic brain tissue of rabbits after middle cerebral artery occlusion

Objective To explore the changes of endostatin （a strong anti-angiogenesis factor） and vascular endothelial growth factor （VEGF） in the brain tissues of rabbits following cerebral ischemia induced by middle cerebral artery occlusion （MCAO）. Methods Twenty-four New Zealand white rabbits were randomly divided into 5 groups： control （n = 5）, sham-operation （n = 4）, 2-hour ischemia （n = 5）, 24-hour ischemia （n = 5）, and 48-hour ischemia （n = 5）. The expression of VEGF and endostatin were measured by enzyme-linked immunosorbent assay （ELISA） and immunohistochemistry, respectively. In situ hybridization was used to characterize the expression of mRNA for the endostatin. Results Both the protein （at least 50%, P 〈 0.01） and mRNA （at least 70%, P 〈 0.05） of endostatin increased significantly in the ischemic brain tissues after MCAO compared with the control group. VEGF increased at least 270% in the brain after cerebral ischemia （P 〈 0.05）. Conclusion Cerebral ischemia leads to an up-regulation of endostatin in the brain, which is not associated with the increase of VEGF in the brain. The increase of endostatin may serve as a deleterious mechanism for ischemic injury through blocking angiogenesis.

晚期结直肠癌的内科治疗

结直肠癌（colorectal cancer,CRC）包括结肠癌和直肠癌,它的发生是一个多因素、多步骤的过程,是由很多因素相互作用的结果,是人类主要的恶性肿瘤之一。在我国其发病率已排到第3位,致死率排第5位,在恶性肿瘤中死亡率已上升到第2位,且随着生活水平的不断提高而不断增加[1]。如果患者能早期诊断,

Effects of Ghrelin Antisense Inhibition on VEGF and Its Receptor Flt-1 mRNA Expression

[ Objective] This study was to investigate the effect of VEGF and its receptor Fit-1 mRNA expression in Mongolia sheep umbilical vein endothelial cells by ghrelin antisense inhibition. [ Method] Experiments were divided into 4 groups： group Ⅰ （blank control group） ; group Ⅱ （liposome group） ; group Ⅲ （SCON group： 20 μmol/L sense oligonucleotide） ; group Ⅳ （ASCON： 20 μmol/L antisense oligonucleotide）. VEGF and its receptor Fit-1 mRNA expression changes were detected by using real-time fluorescence quantitative detection after 24, 36 and 48 h. [ Result] The expression of VEGF mRNA in group Ⅰ, group Ⅱ were insignificantly different at higher expression levels, and did not change significantly with the time; the expression of VEGF mRNA in group Ⅲ assumed a slight decrease, but there were no significant differences between group I and group Ⅱ （P 〉0.05）, the expression of VEGF mRNA in group Ⅳ（antisense oligonucleotide group ） decreased significantly （P 〈 0.05） ; the expression of VEGF receptor FLT-1 mRNA was similar to that of VEGF. [ Conclusion] Antisense inhibition ghrelin has a downward effect to the expression of VEGF and its receptor Fit-1 the mRNA.

硬膜外镇痛与静脉镇痛对子宫肌瘤患者血清CA125、VEGF-C、PGE2的影响

目的:探讨硬膜外镇痛与静脉镇痛对子宫肌瘤患者血清癌抗原125(cancer antigen 125,CA125)、血管内皮生长因子-C(vascular endothelial growth factor-C,VEGF-C)和前列腺素E2(prostaglandin E_2,PGE2)的影响。方法:选自我院于2015年9月至2017年9月期间收治的子宫肌瘤患者98例,按照随机表法分为观察组49例与对照组49例。两组均采用蛛网膜下腔阻滞麻醉。对照组采用自控静脉镇痛,观察组采用硬膜外麻醉。比较两组手术时间和术中出血量,术前和术后VAS评分、CA125、VEGF-C和PGE2水平变化。结果:两组手术时间和术中出血量比较无统计学差异(P>0.05)。两组术后3、24、48hVAS评分较术前降低(P<0.05);观察组术后3、24、48hVAS评分低于同期对照组(P<0.05)。两组术后48h血清CA125、VEGF-C和PGE2水平较术前降低(观察组:t=12.867、16.511、16.674,对照组:t=7.987、7.608、8.425,P<0.05);观察组术后48h血清CA125、VEGFC和PGE2水平低于对照组(t=7.442、10.418、10.682,P<0.05)。结论:硬膜外镇痛对子宫肌瘤效果明显,且可降低CA125、VEGF-C和PGE2水平,具有重要研究意义。

阿芬太尼对糖尿病大鼠心肌缺血再灌注损伤及VEGFA/Ang-1通路的影响

目的:探讨阿芬太尼对糖尿病大鼠心肌缺血/再灌注(I/R)损伤及血管内皮生长因子A(VEGFA)/血管紧张素-1(Ang^(-1))通路的影响。方法:用链脲佐菌素(60 mg·kg^(-1))腹腔注射的方法制备大鼠糖尿病模型,造模成功后制备I/R模型。将造模成功的Wistar大鼠随机分为假手术组、I/R组、吗啡组(0.1 mg·kg^(-1))和阿芬太尼低、高剂量组(3,6 mg·kg^(-1)),每组10只。在结扎前30 min,阿芬太尼低、高剂量组和吗啡组尾静脉注射相应剂量药物,假手术组和I/R组尾静脉注射等量生理盐水。再灌注2 h,对大鼠进行心功能检测,处死大鼠,检测血清中肌酸激酶同工酶(CK-MB)、乳酸脱氢酶(LDH)、丙二醛(MDA)和超氧化物歧化酶(SOD)水平;检测心肌组织中VEGFA、Ang^(-1)和半胱氨酸天冬氨酸蛋白酶-3(caspase-3)蛋白水平。结果:与假手术组比较,其余各组大鼠左心室舒张压(LVDP)、血清中SOD、心肌组织中VEGFA和Ang^(-1)蛋白水平降低,左心室舒张末期内径(LVEDD)、左心室收缩末期内径(LVESD)、血清中CK-MB、LDH、MDA和心肌组织中caspase-3水平升高(P<0.05)。与I/R组比较,阿芬太尼低、高剂量组和吗啡组大鼠LVDP、血清中SOD、心肌组织中VEGFA和Ang^(-1)蛋白水平升高,LVEDD、LVESD、血清中CK-MB、LDH、MDA和心肌组织中caspase-3水平降低(P<0.05)。阿芬太尼低剂量组与高剂量组、吗啡组比较,以上各指标差异有统计学意义(P<0.05);阿芬太尼高剂量组和吗啡组作用相似,差异无统计学意义(P>0.05)。结论:阿芬太尼可以激活VEGFA/Ang^(-1)通路,降低caspase-3水平,对糖尿病大鼠心肌I/R损伤发挥保护作用。

C-reactive protein,procalcitonin,interleukin-6,vascular endothelial growth factor and oxidative metabolites in diagnosis of infection and staging in patients with gastric cancer

AIM:The current study was to determine the serum/pLasma levels of VEGF,IL-6,malondialdehyde (MDA),nitric oxide (NO),PCT and CRP in gastric carcinoma and correlation with the stages of the disease and accompanying infection. METHODS:We examined the levels of serum VEGF,IL-6, PCT,CRP and plasma MDA,NO in 42 preoperative gastric cancer patients and 23 healthy subjects.There were infection anamneses that had no definite origin in 19 cancer patients. RESULTS:The VEGF levels (mean±SD; pg/mL) were 478.05±178.29 and 473.85±131.24 in gastric cancer patients with and without infection,respectively,and these values were not significantly different (P>0.05).The levels of VEGF, CRP,PCT,It-6,MDA and NO in cancer patients were significantly higher than those in healthy controls and the levels of CRP,PCT,It-6,MDA and NO were statistically increased in infection group when compared with non- infection group (P<0.001). CONCLUSION:Although serum VEGF concentrations were increased in gastric cancer,this increase might not be related to infection.CRP,PCT,IL-6,MDA and NO have obvious drawbacks in the diagnosis of infections in cancer patients. These markers may not help to identify infections in the primary evaluation of cancer patients and hence to avoid unnecessary antibiotic treatments as well as hospitalization. According to the results of this study,IL-6,MDA,NO and especially VEGF can be used as useful parameters to diagnose and grade gastric cancer.

Integrin-linked kinase in gastric cancer cell attachment,invasion and tumor growth

AIM： To investigate the effects of integrin-linked kinase （ILK） on gastric cancer cells both in vitro and in vivo. METHODS： ILK small interfering RNA （siRNA） was transfected into human gastric cancer BGC-823 cells and ILK expression was monitored by real-time quan- titative polymerase chain reaction, Western blotting analysis and immunocytochemistry. Cell attachment, proliferation, invasion, microfilament dynamics and the secretion of vascular endothelial growth factor （VEGF） were also measured. Gastric cancer cells treated with ILK siRNA were subcutaneously transplanted into nude mice and tumor growth was assessed. RESULTS： Both ILK mRNA and protein levels were significantly down-regulated by ILK siRNA in human gastric cancer cells. This significantly inhibited cell attachment, proliferation and invasion. The knockdown of ILK also disturbed F-actin assembly and reduced VEGF secretion in conditioned medium by 40% （P 〈 0.05）. Four weeks after injection of ILK siRNA-transfected gastric cancer cells into nude mice, tumor volume and weight were significantly reduced compared with that of tumors induced by cells treated with non-silencing siRNA or by untreated cells （P 〈 0.05）. CONCLUSION： Targeting ILK with siRNA suppresses the growth of gastric cancer cells both in v/tro and /n vivo. ILK plays an important role in gastric cancer progression.

Insulin promotes sinusoidal endothelial cell proliferation mediated by upregulation of vascular endothelial growth factor in regenerating rat liver after partial hepatectomy

AIM： To determine whether insulin could promote sinusoidal endothelial cell （SEC） proliferation mediated by upregulation of vascular endothelial growth factor （VEGF） in regenerating rat liver after partial hepatectomy （PHx）. METHODS： Adult male Sprague-Dawley rats undergoing 70% PHx were injected with insulin （300 MU/kg） or saline via the tail veins every 8 h after surgery for 7 d and killed at 0, 24, 48, 72, 96, 120, 144, and 168 h after surgery. Proliferation of both hepatocytes and SECs was monitored by evaluating the proliferating cell nuclear antigen （PCNA） labeling index （LI）. The expression of VEGF protein was evaluated by immunohistochemistJv. The mRNA expressions of VEGF and its receptors FIt-1 and FIk-1 were evaluated by semi-quantitative reverse transcription-PCR. RESULTS： Insulin markedly increased the expression of VEGF mRNA between 24 and 120 h after hepatectomy compared to controls. Similarly, insulin significantly increased the expression of Fit-1 between 24 and 96 h. However, insulin had no significant effect on FIk-1. Furthermore, the immunohistochemical staining revealed that expression of VEGF protein increased in the insulin groups. Insulin significantly increased the PCNA LI of hepatocytes and SECs compared to controls. CONCLUSION： Exogenous insulin may promote SEC proliferation with an enhanced expression of VEGF and its receptor Fit-1 in regenerating rat liver after PHx.

Curcumin suppresses PPARδ expression and related genes in HT-29 cells

AIM:To investigate the effects of curcumin on the expression of peroxisome proliferator-activated receptorδ(PPARδ)and related genes in HT-29 cells. METHODS:HT-29 cells were treated with curcumin (0-80μmol/L)for 24 h.The effects of curcumin on the morphology of HT-29 cells were studied by Hoechst 33342 staining.The activity of caspase-3 was determined using DEVD-p NA as substrate.The levels of peroxisome PPARδ,14-3-3εand vascular endothelial growth factor(VEGF)in HT-29 cells were determined by Western blotting analysis and their mRNA expression was determined by real-time quantitative RT-PCR. RESULTS:Treatment with 10-80μmol/L curcumin induced typical features of apoptosis and activated the caspase-3 in HT-29 cells.The expression of PPARδ, 14-3-3εand VEGF was reduced and the activity of β-catenin/Tcf-4 signaling was inhibited by curcumin treatment. CONCLUSION:Curcumin can induce apoptosis of HT-29 cells and down-regulate the expression of PPARδ,14-3-3εand VEGF in HT-29.

(-)-Epigallocatechin-3-gallate inhibits VEGF expression induced by IL-6 via Stat3 in gastric cancer

AIM: To demonstrate that (-)-Epigallocatechin-3-gallate (EGCG) inhibits vascular endothelial growth factor (VEGF) expression and angiogenesis induced by interleukin-6 (IL-6) via suppressing signal transducer and activator of transcription 3 (Stat3) activity in gastric cancer. METHODS: Human gastric cancer (AGS) cells were treated with IL-6 (50 ng/mL) and EGCG at different concentrations. VEGF, total Stat3 and activated Stat3 protein levels in the cell lyses were examined by Western blotting, VEGF protein level in the conditionedmedium was measured by enzyme-linked immunosorbent assay, and the level of VEGF mRNA was evaluated by reverse transcription polymerase chain reaction (RTPCR). Stat3 nuclear translocation was determined by Western blotting with nuclear extract, and Stat3-DNA binding activity was examined with Chromatin immunoprecipitation (ChIP) assay. IL-6 induced endothelial cell proliferation was measured with 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl tetrazoliumbromide assay, in vitro angiogenesis was determined with endothelial cell tube formation assay in Matrigel, and IL-6-induced angiogenesis in vitro was measured with Matrigel plug assay. RESULTS: There was a basal expression and secretion of VEGF in AGS cells. After stimulation with IL-6, VEGF expression was apparently up-regulated and a 2.4-fold increase was observed. VEGF secretion in the conditioned medium was also increased by 2.8 folds. When treated with EGCG, VEGF expression and secretion were dose-dependently decreased. IL-6 also increased VEGF mRNA expression by 3.1 folds. EGCG treatment suppressed VEGF mRNA expression in a dose-dependent manner. EGCG dose-dependently inhibited Stat3 activation induced by IL-6, but did not change the total Stat3 expression. When treated with EGCG or AG490, VEGF expressions were reduced to the level or an even lower level in the tumor cells not stimulated with IL-6. However, PD98059 and LY294002 did not change VEGF expression induced by IL-6. EGCG inhibited Stat3 nucleus translocation, and Stat3-DNA binding activity was also markedly decreased by EGCG. Furthermore, EGCG inhibited IL-6 induced vascular endothelial cell proliferation and tube formation in vitro and angiogenesis in vitro . CONCLUSION: EGCG inhibits IL-6-induced VEGF expression and angiogenesis via suppressing Stat3 activity in gastric cancer, which has provided a novel mechanistic insight into the anti-angiogenic activity of EGCG.

REGULATING EFFECTS OF VASCULAR ENDOTHELIAL GROWTH FACTOR AND ANG Ⅱ ON FROG'S PERICARDIAL STOMATA, MESOTHELIUM AND ANGIOGENESIS

To observe the regulating effects of vascular endothelial growth factor (VEGF) and angiotensinⅡ (ANG II) on the frog’s pericardium, lymphatic stomata and angiogenesis so as to reveal their effects and mechanism on the mesothelial permeability, lymphatic stoma regulation and myocardial hypertrophy. Methods. VEGF and ANGⅡ were injected into the frog’s peritoneal cavity so as to examine the changes of the pericardial stromata by using transmission electron microscopy, scanning electron microscopy and computerized imaging analysis. Results. Scattered distributed pericardial stomata were found on the parietal pericardium of the frog with a few sinusoid mesothelial cells, whose blood supply was directly from the cardiac chambers flowing into the trabecular spaces of the myocardium (because there are no blood vessels in the myocardium of the frog). The average diameters of the pericardial stomata in VEGF and ANGⅡ groups were 1.50μ m and 1.79μ m respectively, which were much larger than those in the control group (0.72μ m, P Conclusions. VEGF and ANGⅡ could strongly regulate the pericardial stomata by increasing their numbers and openings with larger diameters and higher distribution density. They could also increase the sinusoid areas with the result of the higher permeability of the pericardium, which clearly indicated that VEGF and ANGⅡ could speed up the material transfer of the pericardial cavity and play an important role in preventing myocardial interstitial edema. Yet there was no strong evidence to show the angiogenesis in the myocardium.

Altered expression of mitochondrial related genes in the native Tibetan placents by mitochondrial cDNA array analysis

Objective： To explore the mechanism of native Tibetan fetuses adaptation to hypoxia, we tried to find the different expression genes about mitochondrial function in the native Tibetan placents. Methods： In this study, the placents of native Tibetan and the high-altitude Hart （ha-Hart） were collected. After the total RNA extraction, the finally synthesized cDNAs were hybridized to mitochondrial array to find the altered expression genes between them. Then, the cytochrome c oxidase 17 （Coxl7）, dynactin 2 （DCTN2, also known as p50）, and vascular endothelial growth factor receptor （VEGFR, also known as KDR） were chosen from the altered expression genes to further verify the array results using the SYBR Green real-time PCR. Because the altered expression genes （such as Cybb and Cox 17） in the array results related to the activities of COXI and COXIV, the placental mitochondria activities of COXI and COXIV were measured to find their changes in the hypoxia. Results： By a standard of≥1.5 or ≤0.67, there were 24 different expressed genes between the native Tibetan and the ha-Han placents, including 3 up-regulated genes and 21 down-regulated genes. These genes were related to energy metabolism, signal transduction, cell proliferation, electron transport, cell adhesion, nucleotide-excision repair. The array results of Cox17, DCTN2 and KDR were further verified by the real-time RT-PCR. Through the mitochondria respiration measurements, the activity of COXI in the native Tibetan placents were higher than that of ha-Han, there was no difference in COXIV activity between them. Conclusion： The altered mitochondrial related genes in the native Tibetan placents may have a role in the high altitude adaptation for fetuses through changing the activity of mitochondrial COX.

The effect of rh-endostatin on micrangium and angiogenic factors in tumor and myocardium tissue

Objective： The aim of this study was to compare effect of rh-endostatin on microvasculature in tumor and myocardium tissue. Methods： Nude mice were randomized into 4 groups, blank control group [did not burden tumor, normalsaline （NS） 100 μL/d], drug control group （did not burden tumor, rh-endostatin 400 μg/d）, model group （mice burdened tumor, NS 100 μL/d） and treatment group （mice burdened tumor, rh-endostatin 400 μg/d）, administration was given during d1-d28. The volume of tumor and the weight of mouse were measured before and after administration. The expression of CD34, MMP-2, MMP-9, HIF-la and VEGF in myocardium and tumor were detected by immunohistochemistry. The structure of vasculature was observed by immunoenzymatic double staining with CD34 and Masson. Results： The tumor volume increase of treatment group （48.18 mm3） was less than the model group （113.80 mm3）, the change of weight was not significant among the four groups. After treated with endotar, the expression of MMP-9 and VEGF in tumor were obviously down-regulated, but the same results was not found in MMP-2, HIF-la of tumor. MVD in tumor of treatment group decreased significantly compared with model group. Proportion of tumor vessels covered by collagen in treatment group increased compared with model group. However, MVD and microvasculature in myocardium did not change significantly. Conclusion： Rh-endostatin can decrease the expression of MMP-9, VEGF and MVD to inhibit growth of tumor and normalize micrangium in tumor but cannot weaken MMPs and MVD of mature micrangium in myocardium.

Effect of grape proanthocyanidins on tumor growth and angiogenesis in H22 liver cancer xenograft model

Objective: The aim of this study was to investigate the effect of grape proanthocyanidins(GPC) on the growth and angiogenesis of hepatocellular carcinoma H22 cells xenograft in mice. Methods: The xenograft model was established using injected subcutaneously H22 cells into the right axilla of the mice. Each group was treated with different doses of GPC and Endostar. All these treatments were maintained for 10 days, and mice were sacrificed. The xenograft tumors in mice were measured. The proliferation activity level of H22 cells was determined by MTT assay, and the levels of vascular endothelial growth factor(VEGF) protein were examined by immunohistochemistry. Results: When treated with 50, 100 and 200 mg/kg of GPC and Endostar, the tumor inhibition rates were 13.17%, 23.37%, 36.15% and 14.71%, respectively. The tumor weight of xenograft was significantly lighter in high GPC group than the control group(P < 0.05). The ODs in GPC groups were 0.835, 0.666 and 0.519, respectively. The absorbances in middle and high GPC groups were statistically significant, compared with control group(P < 0.01). Immunohistochemical technique showed the expression of VEGF of the GPC groups was downregulated significantly compared with the control group(P < 0.01). Conclusion: GPC can inhibit the growth of hepatocellular carcinoma H22 cell xenograft in mice. The inhibition of angiogenesis by the down-regulation of VEGF expression may play a key role in the anti-neoplastic effect of GPC.

Effect of grape seed proanthocyanidins on tumor vasculogenic mimicry in liver cancer xenograft model

Objective: As a novel blood supply pattern, vasculogenic mimicry(VM) has attracted increasingly attention in recent years, which may partly compensate for the absence of feeding and facilitate tumor perfusion. However, anti-angiogenic drugs have little effect on VM. The grape seed proanthocyanidins(GSPs), a kind of promising bioactive phytochemical, has shown anti-carcinogenesis and anti-angiogenic in several tumor models. However, GSPs regulation of VM and its possible mechanisms in a H22 hepatoma carcinoma model remain not clear. The aim of this study was to examine the effects of GSPs on proliferation and VM in a H22 hepatoma carcinoma model and to investigate the underlying mechanism. Methods: Seventy-five mice were divided into the control group and experimental groups treated with different concentration of GSPs. CD34-PAS dual staining was employed to identify the VM structure. The immunohistochemical staining for investigating the expression of VEGF, Eph A2 and MMP-2 protein was performed. Results: Treatment of the H22 model with Endostar(4 mg/kg), 50, 100, 200 mg/kg of the GSPs resulted in 6.87%, 17.81%, 27.43%, 53.52% inhibition in tumor growth, respectively. The mean weight of tumors were significantly lower in GSPs(100 mg/kg) and GSPs(200 mg/kg) groups than in the control group(all P < 0.01). Similarly, compared with the control group, the number of VM channels were significantly reduced in GSPs(100 mg/kg) and GSPs(200 mg/kg) groups(all P < 0.01). Immunohistochemistry showed significant decreases in the expression levels of VEGF, Eph A2 and MMP-2 protein in GSPs(100 mg/kg) and GSPs(200 mg/kg) groups when compared with control group(all P < 0.001). Conclusion: This is the first report providing evidence that GSPs inhibit the VM structure by regulation of the VEGF/Eph A2/MMPs signaling pathway. Therefore, we concluded that GSPs has the potential of being a clinical anti-VM inhibitor.

Effect of acupuncture on the luteal function of rats with embryo implantation dysfunction

Objective To observe the effects of acupuncture on the luteal function of rats with embryo implantation dysfunction, and to explore its mechanism. Methods The early pregnant rats were randomly divided into normal control group （N）, model group （M）, acupoint group （A）, and non-acupoint group （AC）. The model of embryo implantation dysfunction in Group M, A, and AC was established with Mifepristone. For rats in Group A, bilateral ＂Housanli＂ （后三里 ST 36） and ＂Sanyinjiao＂ （三阴交 SP 6） were needled, while the non-acupoints beside the acupoints were needled in Group AC. In this experiment, the serum levels of luteinizing hormone （LH）, estradiol （E2）, and progesterone （P） were detected with radioimmunoassay, the expression of vascular endothelial growth factor （VEGF） in the rat ovarian tissue was detected by using Western-blot. The mRNA expressions of VEGF and luteinizing hormone receptor （LHR） in the ovarian tissue were detected by using RT-PCR. Results The serum levels of LH and P were significantly higher in group A than in group M and AC （all P〈0.05）, showing no statistical significance when compared with group N. The VEGF content, and expressions of VEGF mRNA and LHR mRNA in the ovarian tissue of group A were significantly elevated than those in group M and group AC （all P〈0.05）, showing no statistical significance when compared with group N. Conclusion Needling at ＂Housanli＂ （后三里 ST 36） and ＂Sanyinjiao＂ （三阴交 SP 6） could elevate the serum levels of LH and P of rats with embryo implantation dysfunction, and up-regulate the expression of LHR mRNA, VEGF and its mRNA in the ovarian tissue. It may enhance the luteal function of rats with embryo implantation dysfunction and improve its embryo implantation environment.

Use of cationic microbubbles targeted to P-selectin to improve ultrasound-mediated gene transfection of hVEGF_(165) to the ischemic myocardium

Gene therapies have been applied to the treatment of cardiovascular disease, but their use is limited by the need to deliver them to the right target. We have employed targeted contrast ultrasound-mediated gene transfection （TCUMGT） via ultrasound-targeted microbubble destruction （UTMD） to transfer therapeutic genes to specific anatomic and pathological targets. Phospholipid microbubbles （MBs） with pcDNA3.l-human vascular endothelial growth factor 165 （pcDNA3.I-hVEGFls5） plasmids targeted to P-selectin （MB＋P＋VEGFp） were created by conjugating monoclonal antibodies against P-selectin to the lipid shell. These microbubbles were divided into four groups： microbubble only （MB）, microbubble＋P-selectin （MB＋P）, microbubble＋pcDNA3.l-hVEGF185 plasmid （MB＋VEGFp）, and microbubbie＋ P-selectin＋pcDNA3.1-hVEGF185 piasmid （MB＋P＋VEGFp）. The reverse transcription polymerase chain reaction （RT-PCR） and enzyme-linked immunosorbent assay （ELISA） results showed that the VEGF gene was successfully transfected by TCUMGT and the efficiency is increased with P-selectin targeting moiety. UTMD-mediated delivery of VEGF increased myocardial vascular density and improved cardiac function, and MB＋P＋VEGFp delivery showed greater improvement than MB＋VEGFp. This study drew support from TCUGMT technology and took advantage of targeted ultrasound contrast agent to identify ischemic myocardium, release pcDNA3.1-hVEGF165 recombinant plasmid, and improve the myocardial microenvironment, so promoting the restoration of myocardial function.