Expression of thymidine kinase mediated by a novel non-viral delivery system under the control of vascular endothelial growth factor receptor 2 promoter selectively kills human umbilical vein endothelial cells

AIM: To investigate the killing efficiency of a recombinant plasmid containing a thymidine kinase (TK) domain insert driven by the vascular endothelial growth factor receptor 2 (VEGFR2) promoter (KDR) on vascular endothelial cells.METHODS: The KDR-TK fragment was extracted from pBluescript Ⅱ KDR-TK plasmid by enzymatic digestion with Xho I and Sal I. The enhanced green fluorescence protein (EGFP) carrier was extracted from pEGFP by the same procedure. The KDR-TK was inserted into the pEGFP carrier to construct pEGFP-KDR-TK. Using ultrasound irradiation and microbubble, pEGFP-KDR-TK was transferred into human umbilical vein endothelial cells (HUVECs). The transient infection rate was estimated by green fluorescent protein (GFP) expression. Transfected HUVECs, non-transfected HUVECs, and HepG2 cells were cultured in the presence of different concentrations of ganciclovir (GCV), and the killing efficacy of HSV-TK/GCV was analyzed by 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide (MTT) assay. RESULTS: The recombinant pEGFP-KDR-TK was successfully constructed by inserting the KDR-TK fragment into the pEGFP carrier. Transfected HUVECs showed cytoplasmic green fluorescence, and the transient transfection rate was about 20.3%. Pools of G418-resistant cells exhibited a higher sensitivity to theprodrug/GCV compared to non-transfected HUVECs or non-transfected HepG2 cells, respectively. CONCLUSION: KDR promoter and the suicide gene/prodrug system mediated by diagnostic ultrasound combined with microbubble can significantly kill HUVECs. Such therapy may present a novel and attractive approach to target gene therapy on tumor vessels.

Increased serum levels of soluble CD146 and vascular endothelial growth factor receptor 2 in patients with exudative age-related macular degeneration

AIM: To investigate serum levels of soluble CD146(s CD146) and vascular endothelial growth factor receptor 2(VEGFR2) in patients with age-related macular degeneration(AMD). METHODS: Eighty-eight patients with exudative AMD and 45 sex-and age-matched healthy controls were enrolled in this study conducted in China. Serum samples was obtained from the patients with exudative AMD and from the controls. Serum sCD146 and VEGFR2 protein levels were measured using an enzyme-linked immunosorbent assay.RESULTS: We found that serum sCD146 and VEGFR2 protein levels were significantly higher in the patients with exudative AMD group than in the controls(t=3.859, P<0.001 and t=3.829, P<0.001, respectively). Serum sCD146 levels were significantly higher in patients with classic choroidal neovascularization(CNV) than in those with occult CNV(t=9.899, P<0.001). There was a significant difference in the trend for exudative AMD in the highest versus lowest quartile of circulating sCD146 levels(χ2=10.29, P=0.001). The receiver operating characteristic curve analysis showed that the area under the curve was 0.696 for s CD146(95%CI: 0.601-0.791) with an optimum diagnostic cut-off value of 157.16 ng/mL, a sensitivity of 55.7%, and a specificity of 82.2%.CONCLUSION: The serum sCD146 level increases and may be a biomarker for exudative AMD.

Specific anti-tumor effect induced by attenuated Salmonella typhimurium vaccine expressing extracellular region of vascular endothelial growth factor receptor 2

The purposes of this research were to study the stable expression of exogenous gene encoding therapeutic protein in attenuated Salmonella typhimurium, observe the metabolism of oral gene vaccine carried by attenuated Salmonella typhimurium in BALB/c mouse, and investigate the feasibility of prevention and treatment of tumors by the recombinant bacteria. Recombinant plasmid pcDNA3.1+ VEGFR2(n1-7) was transformed into competent attenuated Salmonella typhimurium SL3261 to develop oral DNA vaccine SL3261-pcDNA3.1+VEGFR2(n1-7). To observe whether the exogenous gene can be expressed in the recombinant bacteria, PCR was performed to amplify the CMV promoter of the eukaryotic expression vector as the proof of stable expression of exogenous protein; transmission elec- tron microscopy (TEM) was applied to observe the morphology of the recombinant bacteria to confirm that the exogenous gene has no impact on the growth of the bacteria, and then BALB/c mice were immunized with the gene vaccine. After inoculation of the gene vaccine, the recombinant bacteria SL3261 could be detected in the tissues such as small intestine, colon, liver and spleen. And then, mice in each group were challenged with tumor cells. The results of animal experiment showed that tumor growth of the mice in experimental group was inhibited and survival time of immunized mice was prolonged compared with control groups. A higher lymphocyte infiltration in tumors from animals treated with DNA vaccine was observed. Immunohistochemical analysis of tumor samples revealed an en- hanced accumulation of CD8+ cytotoxic T lymphocytes, as well as an increase in CD4+ cells in the tumors of animals treated with the oral gene vaccine compared to tumors from control group mice. Ultrastructure of the tumor tissue showed that tumor cells in the samples of the immunized mice were well-differentiated. Our research confirmed that the exogenous gene can be stably expressed in the attenuated Salmonella typhimurium and has no impact on the growth of the recombinant bacteria; the exogenous gene can de delivered to the host by attenuated Salmonella typhimurium to produce anti-tumor effect with no obvious cytotoxity to the host. In this study, it is established that attenuated Salmonella typhimurium could be used as a vector for oral gene vaccine, and our study provided a theoretical basis for the body distribution and the metabolism of the recombinant bacteria. This strategy may provide a simple, safe and effective way for the prevention and treatment of tumors.

Effects of Huatan Tongluo decoction on vascular endothelial growth factor receptor 2 expression in synovial tissues of rats with collagen-induced arthritis

OBJECTIVE: To determine the therapeutic effect and potential mechanism of Huatan Tongluo decoction on rats with collagen-induced arthritis.METHODS: Forty specific pathogen-free Wistar rats were selected, and 10 were randomly selected as the control(group 1). The remaining rats were injected intradermally with emulsified type Ⅱ bovine collagen at the tail base and back, followed by a booster 7 d post first immunization. After establishing collagen-induced arthritis(CIA), rats were randomly divided into three groups(n = 10). The rats were treated orally for 30 d as follows: group 1, saline; group 2, model(saline); group 3, tripterygium polyglycoside(TP; 7.81 mg/kg, positive control);group 4, Huatan Tongluo decoction(HTTL; 7.5 g/kg). Body weight, ankle swelling and arthritis index were measured over the course of the study. The rats were sacrificed 30 d after treatment. Morphological changes in the synovium were observed by hematoxylin and eosin staining. Pannus formation and synovial thickness in the left ankle were observed by color Doppler ultrasoundVascular endothelial growth factor(VEGF) and VEGFR2 protein levels were measured by immunohistochemistry.VEGF/VEGFR2 mRNA levels were measured by real-time quantitative polymerase chain reaction.RESULTS: Compared with the model group, a significantly lower arthritis index was observed in the positive control group(P < 0.05) and HTTL group(P < 0.01), after treatment. Both positive control and HTTL reduced intra-articular pannus formation and synovial thickening. Furthermore, VEGF mRNA,and VEGFR2 protein and mRNA levels were significantly downregulated(P < 0.05) in the treatment groups.CONCLUSION: Inhibition of the expression of VEGF and VEGFR2 in synovial tissues and the formation of pannus and synovial hyperplasia may be part of the mechanism of HTTL for relieving the symptoms of rheumatoid arthritis in CIA rats.

Molecular Modeling Studies of Vascular Endothelial Growth Factor Receptor Tyrosine Kinase Inhibitors Combining Molecular Docking and 3D-QSAR Methods

The vascular endothelial growth factor （VEGF） and its receptor tyrosine kinases VEGFR-2 or kinase insertdomain receptor （KDR） have emerged as attractive targets for the design of novel anticancer agents. In the present work, molecular docking method combined with three dimensional quantitative structure-activity relationships （comparative molecular field analysis （CoMFA） and comparative molecular similarity indice analysis （CoMSIA）） to analyze the possible interactions between KDR and those derivatives which acted as selective inhibitors. The CoMFA and CoMSIA models gave a cross-validated coefficient Q2 of 0.713 and 0.549, non-cross-validated R2 values of 0.974 and 0.878, and predicted R2 values of 0.966 and 0.823, respectively. The 3D contour maps generated by the CoMFA and CoMSIA models were used to identify the key structural requirements responsible for the biological activity. The information obtained from 3D-QSAR and docking studies were very helpful to design novel selective inhibitors of KDR with desired activity and good chemical property.

Serum vascular endothelial growth factor receptor-2 and adropin levels in age-related macular degeneration

AIM： To investigate the serum levels of vascular endothelial growth factor receptor-2（VEGFR-2） and adropin in age-related macular degeneration（AMD）patients.·METHODS： Ninety-eight AMD patients were included in the study. Seventy-eight age- and sex-matched healthy volunteers were recruited as the control group.Fundus florescein angiography and optical coherence tomography were performed to assess the posterior segment details. Serum VEGFR-2 and adropin levels were measured using enzyme-linked immunosorbent assays and compared between the study groups.· RESULTS： AMD group had significantly increased foveal retinal thickness, serum LDL and HDL levels and significantly decreased subfoveal choroidal thickness（P =0.01, 0.047, 0.025 and 〈0.001, respectively）. Serum VEGFR-2level revealed a significant decrease in AMD patients compared to controls（26.48 ±6.44 vs 30.42 ±7.92 ng/m L,P 〈0.001）. There was an insignificant increase in serum adropin level in AMD patients（6.17±3.19 vs 5.79±2.71 ng/m L,P =0.4）. Serum level of VEGFR-2 in AMD patients had a significant negative correlation with foveal retinal thickness（r =-0.226, P =0.025） and a significant positive correlation with subfoveal choroidal thickness（r=0.2, P=0.048）.·CONCLUSION： The current study demonstrated that the decreased serum VEGFR-2 level may be considered in the development of AMD. Adropin does not seem to play a role in the pathogenesis of AMD.

Three-dimensional-arterial spin labeling perfusion correlation with diabetes-associated cognitive dysfunction and vascular endothelial growth factor in type 2 diabetes mellitus rat

BACKGROUND Type 2 diabetes mellitus(T2DM) has been strongly associated with an increased risk of developing cognitive dysfunction and dementia.The mechanisms of diabetes-associated cognitive dysfunction(DACD) have not been fully elucidated to date.Some studies proved lower cerebral blood flow(CBF) in the hippocampus was associated with poor executive function and memory in T2DM.Increasing evidence showed that diabetes leads to abnormal vascular endothelial growth factor(VEGF) expression and CBF changes in humans and animal models.In this study,we hypothesized that DACD was correlated with CBF alteration as measured by three-dimensional(3D) arterial spin labeling(3D-ASL) and VEGF expression in the hippocampus.AIM To assess the correlation between CBF(measured by 3D-ASL and VEGF expression) and DACD in a rat model of T2DM.METHODS Forty Sprague-Dawley male rats were divided into control and T2DM groups.The T2DM group was established by feeding rats a high-fat diet and glucose to induce impaired glucose tolerance and then injecting them with streptozotocin to induce T2DM.Cognitive function was assessed using the Morris water maze experiment.The CBF changes were measured by 3D-ASL magnetic resonance imaging.VEGF expression was determined using immunofluorescence.RESULTS The escape latency time significantly reduced 15 wk after streptozotocin injection in the T2DM group.The total distance traveled was longer in the T2DM group;also,the platform was crossed fewer times.The percentage of distance in the target zone significantly decreased.CBF decreased in the bilateral hippocampus in the T2DM group.No difference was found between the right CBF value and the left CBF value in the T2DM group.The VEGF expression level in the hippocampus was lower in the T2DM group and correlated with the CBF value.The escape latency negatively correlated with the CBF value.The number of rats crossing the platform positively correlated with the CBF value.CONCLUSION Low CBF in the hippocampus and decreased VEGF expression might be crucial in DACD.CBF measured by 3D-ASL might serve as a noninvasive imaging biomarker for cognitive impairment associated with T2DM.

Vascular endothelial growth factor A promotes platelet adhesion to collagen Ⅳ and causes early brain injury after subarachnoid hemorrhage

The role of vascular endothelial growth factor A in platelet adhesion in cerebral microvessels in the early stage of subarachnoid hemorrhage remains unclear.In this study,the endovascular puncture method was used to produce a rat model of subarachnoid hemorrhage.Then,30 minutes later,vascular endothelial growth factor A antagonist anti-vascular endothelial growth factor receptor 2 antibody,10μg,was injected into the right ventricle.Immunohistochemistry and western blot assay were used to assess expression of vascular endothelial growth factor A,occludin and claudin-5.Immunohistochemical double labeling was conducted to examine co-expression of GP Ⅰa-Ⅱ integrin and type Ⅳ collagen.TUNEL was used to detect apoptosis in the hippocampus.Neurological score was used to assess behavioral performance.After subarachnoid hemorrhage,the expression of vascular endothelial growth factor A increased in the hippocampus,while occludin and claudin-5 expression levels decreased.Co-expression of GP Ⅰa-Ⅱ integrin and type Ⅳ collagen and the number of apoptotic cells increased,whereas behavioral performance was markedly impaired.After treatment with anti-vascular endothelial growth factor receptor 2 antibody,occludin and claudin-5 expression recovered,while co-expression of GP Ⅰa-Ⅱ integrin and type Ⅳ collagen and the number of apoptotic cells decreased.Furthermore,behavioral performance improved notably.Our findings suggest that increased vascular endothelial growth factor A levels promote platelet adhesion and contribute to early brain injury after subarachnoid hemorrhage.This study was approved by the Biomedical Ethics Committee,Medical College of Xi’an Jiaotong University,China in December 2015.

VEGFR2、miR-21、UCP1及UCP3在非小细胞肺癌组织中表达及与其预后的相关性分析

目的分析血管内皮细胞生长因子受体2(VEGFR2)、微小RNA-21(miR-21)、解偶联蛋白1(UCP1)、解偶联蛋白3(UCP3)在非小细胞肺癌(NSCLC)组织内的表达及与其预后的相关性。方法选取98例NSCLC患者,术中取其癌组织与癌旁正常组织,检测VEGFR2、miR-21、UCP1及UCP3表达;分析VEGFR2、miR-21、UCP1及UCP3表达与其临床病理特征的关系;随访1年,分析VEGFR2、miR-21、UCP1及UCP3表达与患者生存率的关系。结果癌组织内的VEGFR2、UCP1阳性表达率及miR-21相对表达量高于癌旁正常组织,UCP3阳性表达率低于癌旁正常组织,差异有统计学意义(P<0.05);VEGFR2、miR-21、UCP1及UCP3表达与淋巴结转移、临床分期、分化程度有关(P<0.05);VEGFR2、UCP1阳性表达及miR-21高表达患者的1年生存率分别低于VEGFR2、UCP1阴性表达及miR-21低表达患者,UCP3阳性表达患者的1年生存率高于UCP3阴性表达者,差异有统计学意义(P<0.05)。结论VEGFR2、miR-21、UCP1及UCP3在NSCLC癌组织内呈异常表达,与患者的预后具有紧密联系。

Effect of Kang’ai Injection (康艾注射液) on Serum Level of Soluble Interleukin-2 Receptor and Vascular Endothelial Growth Factor in Patients with Esophageal Carcinoma during Radiotherapy

Objective： To observe the effect of Kang＇ai Injection （康艾注射液, KAI） on serum level of soluble interleukin-2 receptor （sIL-2R） and vascular endothelial growth factor （VEGF） in patients with esophageal carcinoma （EC） during radiotherapy （RT）, and to investigate its synergistic effect with RT and its influence on immunological function of the body. Methods： One hundred and seventy patients with EC, who had missed the chance of surgical operational therapy, were assigned to the treated group （90 cases） and the RT group （80 cases）, and at the same time a control group consisting of 80 inpatients without tumors was set up. Patients in the RT group were treated with RT alone but KAI was given additionally to those in the treated group, with 50 ml given once per day via intravenous dripping, 15 days as one course, and 2 courses administered in total. The immediate therapeutic efficacy and changes of serum slL-2R and VEGF levels were observed, and the effect of KAI on patients＇ quality of life （QOF） was evaluated by Karnofsky scoring. Results： In 16 patients of the treated group it was completely remission （OR）, in 54 partially remission （PR）, in 18 it was stabilized disease （SD） and in 2 progressive disease （PD）, with the total effective rate （CR ＋ PR） as 77.8%, while in those of the control group it was 12, 46, 18, 4 and 72.5%, respectively, the immediate therapeutic efficacy in the treated group was somewhat better than that in the RT group, but showed no statistical significance （P〉0.05）. Serum levels of slL-2R and VEGF in all the patients before treatment were higher than those in the control group, which were decreased after treatment in both groups （P〈0.05）, but the improvement in the treated group was better than that in the RT group, showing significant difference （P〈0.05）, and patients＇ QOF improved more significantly in the former as well （62.2 % vs 40.0%, P〈 0.05）. Conclusion： KAI in combination with RT in treating patients with EC could enhance the immunological function of patients, improve their QOF and enhance their sensitivity to RT.

创面血管内皮生长因子、血管内皮生长因子受体2水平表达与Ⅱ期肛裂患者术后创面愈合时间的关系研究

目的探讨创面血管内皮生长因子(vascular endothelial growth factor,VEGF)、血管内皮生长因子受体(vascular endothelial growth factor receptor,VEGFR)-2表达与Ⅱ期肛裂术后创面愈合时间的关系。方法选取在我院进行手术治疗的Ⅱ期肛裂患者104例为研究对象。术后1周换药时采集创缘肉芽组织检测VEGF、VEGFR-2水平。统计患者术后创面愈合时间,根据患者术后创面愈合时间三分位数分3组,对比3组临床资料及创面VEGF、VEGFR-2水平。分析VEGF、VEGFR-2与术后创面愈合时间的相关性及术后创面愈合时间的影响因素。结果3组创面VEGF及VEGFR-2水平比较,差异有统计学意义(P<0.05);创面VEGF、VEGFR-2水平愈低,其创面愈合时间愈长(P<0.05);VEGF、VEGFR-2与术后创面愈合时间呈负相关(P<0.05);便秘、手术时间、开放切口、创面纵径、创面横径为术后创面愈合时间的独立危险因素,VEGF、VEGFR-2均术后创面愈合时间的独立保护因素(P<0.05)。结论创面VEGF、VEGFR-2水平与Ⅱ期肛裂术后创面愈合时间呈负相关,可通过检测两指标水平判断患者创面愈合情况,为术后治疗方案的完善提供指导。

Efficacy of Ciji Hua'ai Baosheng formula on the expressions of vascular endothelial growth factor,kinase insert domain-containing receptor and basic fibroblast growth factor in mouse models of H22 hepatocellular carcinoma

OBJECTIVE: To investigate the efficacy of Ciji Hua'ai Baosheng formula(CHBF) on microvessel density(MVD) and vascular endothelial growth factor(VEGF), kinase insert domain-containing receptor(KDR) and basic fibroblast growth factor(b FGF) expression in serum and tumor tissue of mice receiving chemotherapy for the treatment of H22 hepatocellular carcinoma.METHODS: Sixty Kunming mice were injected subcutaneously with H22 hepatoma carcinoma cell suspensions into the right anterior armpit. Seven days later, all transplanted tumor were formed and the mice were intraperitoneally injected 200 mg/kg cytoxan(CTX) to establish the models of tumor-bearing mouse chemotherapy, then they were randomly divided into model group, continuing CTX chemotherapy group(CTX group), and three CHBF(117, 58.5 and 29.25 g/kg) groups. After ten days of treatments, histology was observed, contents of VEGF, KDR and b FGF in serum and tumor tissue were measured by enzyme-linked immunosorbent assay(ELISA), VEGF and b FGF protein expression and MVD tagged by CD34 were detected by immunohistochemisty.RESULTS: MVD in CHBF(117, 58.5 g/kg) and CTX groups was significantly lower than that in model group(P < 0.01); expressions of VEGF, KDR and b FGF in serum and tumor tissue in CHBF(117 g/kg)group were less than those in model group(P <0.05; P < 0.01); the expressions of MVD, VEGF and b FGF in tumor tissue of CHBF(117 g/kg) groupwere also less than those in CTX group(P < 0.05;P < 0.01).CONCLUSION: CHBF can effectively reduce the expression of VEGF, KDR and b FGF in serum and tumor tissue, and decrease MVD and delay tumor progression.

Stable transfection of estrogen receptor-alpha suppresses expression of cyclooxygenase-2 and vascular endothelial growth factor-C in MDA-MB-231 breast cancer cells

Background Estrogen receptor （ER）-negative breast cancer cells are more aggressive than ER-positive cells. Elevated levels of cyclooxygenase-2 （COX-2） and vascular endothelial growth factor-C （VEGF-C） expression have been detected in cultured human breast cancer cells and are associated with negative hormone receptor status. In this study, we created ERα stable transfectants in MDA-MB-231 cells to explore the effect of ERα on cell growth and COX-2 and VEGF-C expression.Methods The green fluorescent protein （GFP）-ERα plasmids were stably transfected into ER-negative MDA-MB-231 cells. The proliferation and migration of untransfected MDA-MB-231 cells, ERα-transfected MDA-MB-231 cells and ER-positive MCF-7 cells were determined. The expression of COX-2, and the levels of VEGF-C mRNA and the VEGF-C secretion concentration were assayed in these cell lines.Results The proliferation and migration capacities of ERα-tranfected MDA-MB-231 cells were significantly decreased （P 〈0.05）. The expression of COX-2 was significantly lower in ERa-tranfected MDA-MB-231 cells than in untranfected MDA-MB-231 cells. The mRNA and protein levels of VEGF-C were lower in ERa-tranfected MDA-MB-231 cells than in untransfected MDA-MB-231 cells （P〈0.05）.Conclusions ERα stable transfection inhibits proliferation and migration capacities of MDA-MB-231 cells and decreases expression of COX-2 and VEGF-C. The decreases of proliferation and migration capacities may be related to suppression of COX-2 and VEGF-C expression.

联合多期增强CT纹理分析及血液学指标术前预测胃癌VEGFR2表达状态

目的探讨多期动态增强CT纹理分析参数联合血液学指标术前预测胃癌血管内皮生长因子受体2(vascu‐lar endothelial growth factor receptor2,VEGFR2)表达状态的价值。方法选取本院148例胃癌患者资料,获得术前血液学指标和三期增强CT纹理分析参数。基于组内相关系数和差异性检验对参数进行特征筛选。基于二元Logistic回归构建血液学模型、CT纹理分析模型及综合模型来预测VEGFR2表达状态。通过受试者工作特征曲线评估三个模型的诊断效能,并通过列线图来可视化地预测胃癌患者VEGFR2的表达状态。结果基于血液学指标构建的血液学模型曲线下面积(area under the curve,AUC)为0.687。由静脉期纹理分析参数构建的CT纹理分析模型的AUC值为0.624。联合血液学模型和CT纹理分析模型构建的综合模型AUC值为0.723。结论联合多期动态增强CT纹理分析参数及血液学指标有助于术前预测胃癌VEGFR2表达状态。

蒙药香青兰总黄酮对大鼠脑缺血再灌注后缺血半暗带区血管内皮生长因子受体2表达的影响

目的:观察蒙药香青兰总黄酮(total flavones of dracocephalum moldavica,TFDM)对大鼠脑缺血再灌注损伤后缺血半暗带区血管内皮生长因子2(vascular endothelial growth factor 2,VEGFR2)表达的影响,探讨其对血管生成的作用。方法:SD大鼠线栓法制备大脑中动脉闭塞再灌注模型,TFDM 50 mg/kg灌胃给药,在手术后第1、3、7、14天4个时相进行取材,免疫组织化学染色观测VEGFR2阳性标记的细胞及新生血管在缺血半暗带区的分布情况,Western blot检测缺血半暗带区VEGFR2的表达情况。结果:免疫组织化学染色结果显示,再灌注术后第3~14天时,TFDM给药组阳性细胞表达数量显著高于模型组(P<0.05),且在微血管、微动脉和微静脉附近表达增多;Western blot检测结果也显示,TFDM给药组VEGFR2表达上调。结论:TFDM可能通过上调VEGFR2蛋白的表达,促进脑缺血半暗带区微血管的生成,改善微循环功能。

p-AKT与VEGF、HER-2在膀胱浸润性尿路上皮癌中的表达及其与患者预后关系研究

目的探讨磷酸化蛋白激酶B(p-AKT)与血管内皮生长因子(VEGF)、人表皮生长因子受体2(HER-2)在膀胱浸润性尿路上皮癌中的表达及其与患者预后的关系。方法用免疫组织化学染色(IHC)和荧光原位杂交(FISH)检测69例膀胱浸润性尿路上皮癌、20例膀胱黏膜良性组织(慢性炎症、乳头状瘤组织)中p-AKT、VEGF、HER-2蛋白的表达。结果膀胱浸润性尿路上皮癌中p-AKT、VEGF、HER-2表达均显著高于膀胱良性组织的表达(P<0.05);p-AKT、VEGF和HER-2在高级别浸润性尿路上皮癌中的表达均显著高于低级别浸润性尿路上皮癌中的表达(P<0.05);p-AKT在后期复发的患者中的表达高于未复发患者(P<0.05);浸润性尿路上皮癌患者中p-AKT与VEGF、HER-2的表达均呈显著正相关(P<0.05);p-AKT、VEGF和HER-2蛋白表达阳性的患者生存率均明显低于阴性表达患者的生存率(P<0.05)。结论p-AKT、VEGF及HER-2蛋白均可能参与了膀胱浸润性尿路上皮癌的发生发展,并且与患者的预后相关,可能成为膀胱尿路上皮癌的预后指标;膀胱浸润性尿路上皮癌中p-AKT与VEGF、HER-2可能存在相互调节关系。

川楝素调控miR-409-3p对前列腺癌细胞增殖、迁移、侵袭及VEGF/VEGFR2通路的影响

目的探讨川楝素(TSN)调控微小RNA-409-3p(miR-409-3p)对前列腺癌细胞增殖、迁移、侵袭及血管内皮生长因子(VEGF)/血管内皮生长因子受体2(VEGFR2)通路的影响。方法使用0、10、20、40、80和160μmol/L的TSN处理PC3细胞,采用噻唑蓝法检测PC3细胞存活率,选择最佳药物浓度。将细胞分为对照组(Control组)、TSN低浓度组(TSN-L组)、TSN中浓度组(TSN-M组)、TSN高浓度组(TSN-H组)、TSN高浓度+miR-409-3p拮抗剂阴性对照组(TSN-H+antagomiR-NC组)、TSN高浓度+miR-409-3p拮抗剂组(TSN-H+antagomiR-409-3p组)。采用实时荧光定量聚合酶链反应(qRT-PCR)检测PC3细胞中miR-409-3p表达;EdU法检测PC3细胞增殖水平;Transwell小室实验和划痕愈合实验检测PC3细胞的侵袭和迁移能力;Western blot检测细胞周期蛋白D1(CyclinDl)、细胞周期蛋白依赖性激酶4(CDK4)、VEGF、VEGFR2蛋白表达。结果与0μmol/L比较,10、20、40、80和160μmol/L的TSN细胞存活率显著降低(P<0.05),选择20、40、80μmol/L的TSN用于后续实验。与Control组比较,TSN-L组、TSN-M组和TSN-H组PC3细胞EdU阳性率、细胞侵袭数、划痕愈合率、CyclinDl、CDK4、VEGF、VEGFR2蛋白表达显著降低(P<0.05),miR-409-3p表达显著增加(P<0.05),且呈浓度依赖性。与TSN-H+antagomiR-NC组比较,TSN-H+antagomiR-409-3p组PC3细胞EdU阳性率、细胞侵袭数、划痕愈合率、CyclinDl、CDK4、VEGF、VEGFR2蛋白表达显著增加(P<0.05),miR-409-3p表达显著降低(P<0.05)。结论TSN通过上调miR-409-3p表达抑制VEGF/VEGFR2通路的激活,进而抑制PC3细胞增殖、迁移、侵袭。

精神分裂症患者治疗前后的Tie-2、VEGF、PRL变化及其临床意义

目的探讨精神分裂症患者治疗前后血管生成素受体酪氨酸激酶2(Tie-2)、血管内皮生长因子(VEGF)、泌乳素(PRL)的变化及其临床意义。方法回顾性分析2021年7月至2023年9月河南省荣康医院收治的91例精神分裂症患者的临床资料,根据治疗应答情况分为有效组77例和无效组14例。比较两组患者治疗前、治疗2周和4周后的Tie-2、VEGF、PRL水平。应用皮尔逊(Pearson)相关性分析Tie-2、VEGF、PRL与帕利哌酮血药浓度的相关性及其与治疗应答的相关性,绘制受试者工作特征曲线(ROC)评价Tie-2、VEGF及两者联合预测精神分裂症患者治疗后疗效为有效的价值,采用精准-召回曲线评价ROC分析的准确度。结果91例精神分裂症患者治疗后临床痊愈19例,显效48例,好转10例,无效14例,有效率为84.62%;帕利哌酮血药浓度第1~2周呈明显升高趋势,第2周达峰值,第2~4周处于较稳定水平;有效组患者治疗2周和4周后的Tie-2分别为(2153.42±157.83)pg/mL、(2279.88±135.25)pg/mL,明显高于无效组的(1782.19±208.54)pg/mL、(1788.15±223.48)pg/mL,VEGF分别为(422.39±41.65)pg/mL、(461.37±52.80)pg/mL,明显高于无效组的(310.55±24.78)pg/mL、(312.67±27.99)pg/mL,差异均有统计学意义(P<0.05);经Pearson相关性分析结果显示,PRL治疗2周和4周后的变化值与帕利哌酮血药浓度呈显著正相关(r=0.712、0.665,P<0.01);Tie-2和VEGF治疗2周和4周后的变化值与PANSS减分率呈显著正相关(r=0.769、0.752;0.778、0.758,P<0.01);经ROC分析结果显示,Tie-2联合VEGF预测治疗应答的ROC下面积(AUC)为0.899,大于Tie-2(0.769)、VEGF(0.707)(P<0.05);经精准-召回曲线分析结果显示,其AUC为0.901,意味着高精准率和高召回率,采用Tie-2联合VEGF预测治疗应答能为临床医生提供决策支持。结论精神分裂症患者治疗后Tie-2、VEGF变化与帕利哌酮治疗应答有关,联合检测两者可作为预测治疗应答一个方案,为临床医生提供决策支持,PRL变化则与帕利哌酮血药浓度有关,呈现出评估血药浓度和PRL相关不良反应风险的双重应用价值。

Mfn2调控VEGFR2/PI3K促进卵巢癌种植转移的机制研究

目的 探讨线粒体融合蛋白2(Mfn2)对卵巢癌种植转移的作用以及其可能的分子机制。方法 选取2019年6月至2021年6月于南京医科大学附属苏州医院就诊治疗的86例卵巢癌患者作为研究对象,对比癌组织和癌旁组织Mfn2、血管内皮生长因子受体2(VEGFR2)、磷酸酰肌醇3激酶(PI3K)蛋白表达,分析Mfn2、VEGFR2、PI3K蛋白表达与相关病理特征的关系。构建Mfn2上调/下调卵巢癌SKOV-3细胞株并验证Mfn2、VEGFR2、PI3K的表达。结果 Mfn2、VEGFR2、PI3K在肿瘤组织中的阳性表达率高于癌旁组织,差异均有统计学意义(χ^(2)=4.597、6.456、3.930,P=0.032、0.011、0.047);不同FIGO分期、淋巴结转移、远处转移情况及生存情况中,卵巢组织中Mfn2、VEGFR2、PI3K蛋白表达比较,差异均有统计学意义(P<0.05)。与NC组对比,Mfn2上调组卵巢癌SKOV-3细胞中Mfn2 mRNA相对表达量和Mfn2、VEGFR2、PI3K蛋白、显著上调(P<0.05);与shNC组对比,shMfn2下调组卵巢癌SKOV-3细胞中Mfn2mRNA相对表达量和Mfn2、VEGFR2、PI3K蛋白显著下调(P<0.05)。结论 下调Mfn2表达与VEGFR2、PI3K表达水平可以预防卵巢癌种植转移。

清渣汤对玫瑰痤疮模型小鼠皮损组织VEGF、TLR2、KLK5表达的影响

目的:观察清渣汤口服对玫瑰痤疮模型小鼠皮损组织血管内皮生长因子(VEGF)、Toll样受体2(TLR2)、激肽释放酶5(KLK5)表达的影响。方法:7周龄健康雄性小鼠36只,用随机数字表法将其分为6组:清渣汤高剂组、中剂组、低剂组、阳性对照组(多西环素片)、模型对照组、正常对照组,每组6只。除正常组外,其余各组均进行玫瑰痤疮造模。造模24 h后给药,模型组给予0.2 mL生理盐水灌胃,清渣汤3剂组分别给予25 mg/kg(低剂量组)、50 mg/kg(中剂量组)和100 mg/kg(高剂量组)清渣汤灌胃,阳性对照组给予30 mg/kg盐酸多西环素溶液灌胃,每日1次,共给药4周。免疫组化法检测皮损部位皮肤组织VEGF、TLR2、KLK5蛋白表达水平。结果:模型组小鼠皮损组织VEGF、TLR2、KLK5蛋白表达较正常组明显增强(P<0.01);与模型组比较,阳性对照组和清渣汤高、中、低剂量组小鼠皮损组织VEGF、TLR2、KLK5蛋白表达减弱(P<0.05)。结论:清渣汤能够抑制玫瑰痤疮小鼠模型皮损部VEGF、TLR2、KLK5蛋白表达水平,从而减轻炎症反应。