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.

Effects of endostatin on expression of vascular endothelial growth factor and its receptors and neovascularization in colonic carcinoma implanted in nude mice

AIM:To investigate the antiangiogenic effects of endostatin on colonic carcinoma cell line implanted in nude mice and its mechanism. METHODS:Nude mice underwent subcutaneous injection with LS-174t colonic carcinoma cell line to generate carcinoma and were randomly separated into two groups.Mice received injection of vehicle or endostatin every day for two weeks. After the tumor was harvested,the tumor volumes were determined,and the expressions of CD34,VEGF and FIk-1 were examined by immunohistochemical method. RESULTS:Tumor volume was significantly inhibited in the endostatin group(84.17%)and tumor weight was significantly inhibited in the endostatin group(0.197±0.049) compared to the control group(1.198±0.105)(F=22.56, P=0.001),microvessel density(MVD)was significantly decreased in the treated group(31.857±3.515)compared to the control group(100.143±4.290)(F=151.62,P<0.001). Furthermore,the expression of FIk-1 was significantly inhibited in the treated group(34.29%) ompared to the control group(8.57%)(X^2=13.745,P=0.001).However no significant decrease was observed in the expression of vascular endothelial growth factor(VEGF)between these two groups(X^2=0.119,P=0.730). CONCLUSION:Endostatin can inhibit tumor growth and angiogenesis by blocking Vegf/FIk-1 pathway.This experiment provides the theory basis for developing a new anti-carcinoma drug through studying the properties of anti-angiogenesis inhibitors.

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.

Emodin suppresses alkali burn-induced corneal inflammation and neovascularization by the vascular endothelial growth factor receptor 2 signaling pathway

OBJECTIVE:To investigate the effects of emodin on alkali burn-induced corneal inflammation and neovascularization.METHODS:The ability of emodin to target vascular endothelial growth factor receptor 2(VEGFR2)was predicted by molecular docking.The effects of emodin on the invasion,migration,and proliferation of human umbilical vein endothelial cells(HUVEC)were determined by cell counting kit-8,Transwell,and tube formation assays.Analysis of apoptosis was performed by flow cytometry.CD31 levels were examined by immunofluorescence.The abundance and phosphorylation state of VEGFR2,protein kinase B(Akt),signal transducer and activator of transcription 3(STAT3),and P38 were examined by immunoblot analysis.Corneal alkali burn was performed on 40 mice.Animals were divided randomly into two groups,and the alkali-burned eyes were then treated with drops of either 10μM emodin or phosphate buffered saline(PBS)four times a day.Slitlamp microscopy was used to evaluate inflammation and corneal neovascularization(CNV)in all eyes on Days 0,7,10,and 14.The mice were killed humanely 14 d after the alkali burn,and their corneas were removed and preserved at-80℃ until histological study or protein extraction.RESULTS:Molecular docking confirmed that emodin was able to target VEGFR2.The findings revealed that emodin decreased the invasion,migration,angiogenesis,and proliferation of HUVEC in a dose-dependent manner.In mice,emodin suppressed corneal inflammatory cell infiltration and inhibited the development of corneal neovascularization induced by alkali burn.Compared to those of the PBS-treated group,lower VEGFR2 expression and CD31 levels were found in the emodintreated group.Emodin dramatically decreased the expression of VEGFR2,p-VEGFR2,p-Akt,p-STAT3,and p-P38 in VEGF-treated HUVEC.CONCLUSION:This study provides a new avenue for evaluating the molecular mechanisms underlying corneal inflammation and neovascularization.Emodin might be a promising new therapeutic option for corneal alkali burns.

Vitamin D-Binding Protein is Involved in the Pathogenesis of Preeclampsia by Inhibiting the Tyrosine Phosphorylation of Vascular Endothelial Growth Factor Receptor-2 in Endothelial Cells

Objective::The role of Vitamin D-binding protein(DBP)in preeclampsia(PE)pathogenesis is unknown.In this study,we compared the expression of DBP in the placentas of PE patients with the placentas of normotensive pregnant women with placenta previa controls,and aimed to explore the effect of DBP on endothelial cells(ECs)and the underlying mechanism.Methods::DBP expression in placental tissues collected from PE patients and controls was evaluated by immunohistochemistry.The downregulation and upregulation of DBP expression in HTR-8/SVneo cells were examined using DBP-targeting small interfering RNA(siRNA)and DBP-expression vector,respectively.The conditioned media of these DBP-overexpressing and DBP-siRNA HTR-8/SVneo cells were collected and added to human umbilical vein EC(HUVEC)cultures.Angiogenic effects on HUVECs were assessed by tube formation assays,and the proliferation and migration of HUVECs were examined using the Real-Time Cell Analyzer.The expression of vascular endothelial growth factor(VEGF)and VEGF receptor(VEGFR)-2,as well as the phosphorylation of different residues of VEGFR-2 in HUVECs,were determined by western blotting.Results::DBP expression was significantly increased in the placental tissues collected from PE patients.The conditioned medium of DBP-overexpressing HTR-8/SVneo cells potently inhibited tube formation by HUVECs,in addition to their proliferation and migration.Furthermore,treatment of HUVECs with the conditioned medium of DBP-overexpressing HTR-8/SVneo cells decreased the phosphorylation of VEGFR-2 at tyrosine 996,whereas the treatment of these cells with the conditioned medium of DBP-siRNA HTR-8/SVneo cells increased the phosphorylation of VEGFR-2 at tyrosine 951,996,and 1,175.Conclusions::The expression of DBP is increased in the placentas of PE patients.DBP plays potential roles in endothelial dysfunction,which contributes to PE development,by inhibiting tyrosine phosphorylation of VEGFR-2 in ECs.

子宫内膜异位症患者异位内膜中COX-2、VEGF和KDR的表达及意义

目的 探讨环氧化酶-2（COX-2）、血管内皮生长因子（VEGF）和血管内皮生长因子受体（KDR）在子宫内膜异位症（EMs）患者异位内膜组织中的表达。方法 采用免疫组织化学方法测定EMs异位内膜38例（实验组）和正常子宫内膜40例（对照组）中COX-2、VEGF和KDR的表达。结果 ①EMs异位内膜组织中COX-2、VEGF和KDR的表达均较对照组正常子宫内膜的表达高，差异均有统计学意义（P〈0．05）。②在EMs异位内膜组织中，COX-2和VEGF的表达水平呈高度正相关（r=0．984，P〈0．05）。结论异位内膜组织的侵袭力增强及血管生成可能与COX-2、VEGF高表达有关，二者在EMs的发生发展中发挥作用。

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.

胰腺癌p16、Bcl-2、Bax和VEGF基因表达及新生血管密度计数的意义

目的 :研究胰腺癌组织中 p16、 Bcl- 2、 Bax和 VEGF基因的表达及新生血管计数的意义。方法 :采用免疫组织化学法及形态半定量方法对 33例胰腺癌和 6例正常胰腺组织进行新生血管密度记数和 p16、Bcl- 2、Bax和 VEGF基因表达检测。结果 :P16蛋白表达 :高、中分化胰腺癌高于低分化 (P<0 .0 5 ) ,生存期长者表达高于生存期短者 (P<0 .0 5 ) ;Bcl- 2表达 :高、中分化者高于低分化者 ;Bax表达 :正常胰腺组织高于胰腺瘤组织 ,高、中分化者高于低分化者 ,生存期≥ 12个月者高于 <12个月者 ;VEGF表达 :有淋巴结转移者高于无淋巴结转移者 ;MVD记数 :正常胰腺组织低于胰腺瘤组织 ,有淋巴转移者高于无淋巴结转移者 ;MVD与 VEGF表达呈正相关(r=0 .79,P<0 .0 5 ) ;p16基因表达与 VEGF表达和 MVD呈负相关 (r=- 0 .6 9,P<0 .0 5 ;r=- 0 .87,P<0 .0 5 ) ,Bcl- 2表达与其他基因表达结果无相关关系。 Bax表达与 p16表达呈正相关 (r=0 .5 4 ,P<0 .0 5 ) ,与 VEGF表达和 MVD呈负相关 (r=- 0 .6 6 ,P<0 .0 5 )。结论 :综合检测胰腺癌组织中 p16、Bcl- 2、Bax和 VEGF基因的表达及微血管密度对判断胰腺癌恶性程度及估计肿瘤的预后有较大帮助。

具有血管内皮细胞生长因子受体-2酪氨酸激酶抑制作用的链霉菌次生代谢产物2754R的研究

目的分离鉴定链霉菌I06A-02754发酵液中具血管内皮生长因子受体-2酪氨酸激酶(VEGFR2-CD)抑制活性的强极性次生代谢产物。方法采用大孔吸附树脂、阴离子交换树脂、MPLC、HPLC等分离手段对次生代谢产物进行分离纯化;通过UV、IR、HR-ESI质谱、1D-NMR和2D-NMR对其结构进行鉴定,以ELISA法检测其次生代谢产物对VEGFR2-CD的抑制活性;以MTT法检测化合物对肿瘤细胞的抑制活性。结果从发酵液的水溶性部分分离得到一个极性较大的胡桃霉素类次生代谢产物——2754R;其化学结构与胡桃霉素D一致,对VEGFR2-CD表现出一定的抑制活性;MTT实验显示化合物2754R对HepG2细胞、MCF-7细胞和BEL-7402细胞没有明显的抑制活性(IC50>10μmol/L)。结论化合物2754R是具有VEGFR2-CD活性的胡桃霉素类次生代谢产物。

VEGF121和BMP2双基因共表达重组腺病毒载体的构建及其在HEK293中的表达

目的构建人血管内皮生长因子121(VEGF121)与人骨形态发生蛋白2(BMP2)双基因共表达腺病毒载体Adv-BMP2-IRES-VEGF121,并观察其在人胚肾细胞株(HEK293)中的表达情况。方法对腺病毒质粒pShuttle-CMV-BMP2的目的基因BMP2进行PCR扩增。腺病毒质粒pShuttle-CMV-VEGF121-IRES-hrGFP-1经Kpn I/Xba I酶切后,将BMP2片段定向导入pShuttle-CMV-VEGF121-IRES,构建pShuttle-CMV-V EGF121-IRES-BMP2,并注入大肠杆菌DH5a中扩增,提取质粒。通过酶切分析、PCR检测和序列分析进行鉴定。将构建所得的质粒转染HEK293,采用RT-PCR法检测HEK293中的BMP2、VEGF121 mRNA,Western blot法检测其蛋白。结果成功构建了Adv-BMP2-IRES-VEGF121。酶切分析及DNA序列测定证实重组质粒构建正确。质粒转染后的HEK293 BMP2和VEGF121表达阳性。结论成功构建了Adv-BMP2-IRES-VEGF121,其转染HEK293后,VEGF121、BMP2在HEK293中共表达阳性。

KDR mRNA在子宫内膜癌组织中的表达及临床意义

目的研究KDRmRNA的表达与子宫内膜癌发生、发展及生物学行为的关系。方法应用逆转录聚合酶链式反应(RT PCR)技术检测子宫内膜癌45例、子宫内膜不典型增生15例和正常子宫内膜15例组织中的KDRmRNA表达情况。结果KDRmRNA在子宫内膜癌、子宫内膜不典型增生和正常子宫内膜组织中的表达率分别为73.34%(33/45)、26.67%(4/15)和20.00%(3/15),相对含量分别为118.08±15.68、97.59±10.08和80.25±21.38。子宫内膜癌组的KDRmRNA相对含量高于其他两组,P值分别为0.009和0.007;不典型增生组高于正常内膜组,但经比较差异无统计学意义,P=0.431。手术病理分期Ⅲ、Ⅳ期及有淋巴结转移、低分化组的子宫内膜癌KDRmRNA相对含量高于手术病理分期Ⅰ期、无淋巴结转移和高分化组,P值分别为0.012、0.039和0.912;而与肌层浸润深度及病理类型无关,P值分别为0.889和0.912。结论KDRmRNA的过表达在子宫内膜癌的发生、发展中起重要作用,并与子宫内膜癌的生物学行为密切相关,可作为对子宫内膜癌进行治疗的靶位点。

血管内皮生长因子受体2反义核酸体内抑制人乳腺癌生长作用研究

目的探讨血管内皮生长因子受体2(VEGFR2/KDR)反义寡核苷酸(asONs)在体内对人乳腺癌生长的抑制作用。方法构建MCF-7乳腺癌裸鼠模型,将14只裸鼠随机分为生理盐水对照组及反义核酸(asON4)组,asON4按50 mg/kg剂量腹腔注射,1次/d,共20 d;对照组给予相应体积的生理盐水。给药过程中测量各组裸鼠瘤结节的大小;21 d时收集肿瘤标本,免疫组化法检测移植瘤的增殖细胞核抗原(PCNA)表达水平。结果在荷瘤鼠模型中,KDR反义核酸组瘤结节生长变慢,瘤结节大小及移植瘤PCNA阳性细胞数明显低于生理盐水对照组(P<0.05)。结论KDR反义核酸显著抑制KDR蛋白表达,在体内抑制了乳腺癌增殖。

VEGFR 2基因多态性与复发性自然流产的关联性研究

目的探讨血管内皮生长因子受体2(vascular endothelial growth factor receptor 2,VEGFR2/KDR)基因-604T/C,+1192G/A,+1719A/T三个单核苷酸多态性位点与复发性自然流产(recurrent spontaneous abortion,RSA)的关联性。方法采用聚合酶链式反应-限制性片段长度多态性(PCR-RFLP)检测RSA组(180例,病例组)和正常对照组(191例)的VEGFR2基因3个SNPs位点基因型及等位频率分布情况。结果 VEGFR2-604T/C,+1192G/A,+1719A/T的基因型频率在病例组和对照组之间没有统计学意义(P>0.05)。VEGFR2基因+1192G/A位点等位基因分布频率在病例组和对照组间有统计学意义(P<0.01);VEGFR2基因-604T/C,+1192G/A,+1719A/T三个多态性位点所构建的TAA单倍型在病例组和正常对照组间差异有统计学意义(P<0.01)。结论携带VEGFR2+1192G等位基因的基因型可能会增加宁夏汉族女RSA的发病风险,VEGFR2-604/+1192/+1719构建的单倍型TAA与宁夏地区复发性自然流产的发病可能有关。

肺癌组织中KDR基因的表达及其临床意义探讨

目的 :研究KDR (kinasedomain containingreceptor)在肺癌中的表达及临床意义。方法 :采用免疫组化方法 (LSAB法 ) ,检测 114例肺癌组织、癌旁组织 ,3 0例肺良性病变组织中的KDR表达水平。结果 :肺癌组织 ( 86 3 7± 10 90 )中KDR表达水平显著高于肺癌旁组织 ( 4 5 15±2 1 97)和肺良性病变组织 ( 2 0 0 2±11 60 ) ,P <0 0 1,肺癌旁组织显著高于肺良性组织 ,P <0 0 5 ;KDR表达水平与肺癌原发肿瘤大小、淋巴结转移状态 ,P TNM分期和细胞分化程度均有密切关系 ,P <0 0 1,KDR高表达组患者 5年生存率( 2 1 19% )显著低于低表达组 ( 5 2 74% ) ,P <0 0 5。结论 :KDR参与肺癌的发生、发展、转移 。

中国人群VEGFR2 rs2071559与胶质瘤风险的潜在关联性研究

目的探讨中国人群血管内皮细胞生长因子受体2（VEGFR2）多态性与胶质瘤发生风险之间的潜在关联性。方法采用病例-对照研究方法,对504例胶质瘤患者和527例对照者进行血液样本和病历资料收集,以及流行病学问卷调查。Qiagen Blood试剂盒行DNA提取和浓度标化、分装,Mass ARRAY系统等位基因特异性基质辅助激光解析离子化-时间飞跃质谱法完成VEGFR2 rs2071559基因分型,Haplo View 4.1统计软件检验Hardy-Weinberg遗传平衡状态,SPSS 17.0统计软件分析单核苷酸多态性差异。结果 （1）单核苷酸多态性基因分型,VEGFR2 rs2071559分型成功率达99.70%,Hardy-Weinberg遗传平衡检验表明对照组VEGFR2 rs2071559基因型频率处于平衡状态（P=0.451）,具有群体代表性。（2）等位基因频率分析,VEGFR2 rs2071559之C等位基因是胶质瘤危险等位基因,与胶质瘤风险增加有关（OR=1.424,95%CI：1.186~1.710;P=0.000）。（3）基因型分析,VEGFR2 rs2071559之CT和CC基因型与胶质瘤风险增加有关（校正OR=1.407,95%CI：1.071~1.847,P=0.014;校正OR=1.947,95%CI：1.294~2.928,P=0.001）。结论中国人群VEGFR2 rs2071559之CT和CC基因型或C等位基因与胶质瘤风险增加有关,但VEGFR2 rs2071559多态性在胶质瘤易感性中的作用尚待进一步研究。

血管内皮生长因子3’非翻译区936C/T突变与糖尿病视网膜病变的关系

目的:探讨血管内皮生长因子(VEGF)936C/T突变与糖尿病视网膜病变(DR)的关系。方法:按WHO的糖尿病(DM)诊断和排除标准及分型标准,选取无亲缘关系的2型糖尿病患者254例,分为非增殖性视网膜病变(NPDR)组、增殖性视网膜病变(PDR)组和单纯2型糖尿病(DM)组,选取健康体检人群120例作为正常对照组(NC)。采用PCR-RFLP方法确定全部人员的基因型,对不同组间的临床与生化指标及VEGF浓度、936C/T多态性进行了统计分析。结果:NPDR和PDR组的CC基因型频率及C等位基因频率显著高于DM组(2=7.490,2=4.448,P<0.05;2=8.333,2=5.227,P<0.05)和NC组(2=9.934,2=4.899,P<0.05;2=10.895,2=5.714,P<0.05),而NPDR和PDR组(CT+TT)基因型频率及T等位基因频率显著低于NC组(2=9.934,2=10.895,P<0.01;2=4.899,2=5.714,P<0.05)和DM组(2=7.490,2=8.333,P<0.01;2=4.448,2=5.227,P<0.05)。DR多重危险因素分析显示血清低密度脂蛋白胆固醇(LDL-C)、总胆固醇(TC)、糖化血红蛋白(HbA1c)水平以及VEGF浓度和DR发病呈正相关,而VEGF936C/T突变与DR发病危险呈负相关(β=-1.027,OR=0.343,P<0.01,CI:0.157-0.723)。结论:中国汉族人群中存在VEGF936C/T突变。VEGF936C等位基因及CC基因型可能是中国汉族糖尿病患者易于发生DR的危险性遗传标志,而VEGF936T等位基因和携带T等位基因的基因型(936TT基因型和936CT基因型)可能是DR发病风险降低的遗传标志。血浆VEGF、LDL-C、TC和HbA1c水平可能是2型糖尿病患者易发DR的危险因素。

Extraordinary response of metastatic pancreatic cancer to apatinib after failed chemotherapy: A case report and literature review

Chemotherapy has limited efficacy in the treatment of advanced and metastatic pancreatic cancer(PC), and has serious side effects. The development of novel effective agents, especially targeted therapy, is essential for patients with PC. We present a 58-year-old Chinese woman initially diagnosed with locally advanced PC. As the disease progressed to Stage Ⅳ, the patient was unable to tolerate chemotherapy after the fourth-line treatment. She was then treated with apatinib, a novel and highly selective tyrosine kinase inhibitor of vascular endothelial growth factor receptor-2 and achieved a progression-free-survival of 7 mo. All drug-related side effects were well controlled with medication. To the best of our knowledge, this is the first case of PC which responded to apatinib. Considering this remarkable response, apatinib may be a promising agent in the treatment of PC. We also reviewed the literature on chemotherapy and targeted therapy, especially the anti-angiogenesis therapy for patients with PC, and investigated the effect of apatinib in other solid tumors as well.

Ischemic preconditioning protects against ischemic brain injury

In this study, we hypothesized that an increase in integrin αβand its co-activator vascular endothelial growth factor play important neuroprotective roles in ischemic injury. We performed ischemic preconditioning with bilateral common carotid artery occlusion for 5 minutes in C57BL/6J mice. This was followed by ischemic injury with bilateral common carotid artery occlusion for 30 minutes. The time interval between ischemic preconditioning and lethal ischemia was 48 hours. Histopathological analysis showed that ischemic preconditioning substantially diminished damage to neurons in the hippocampus 7 days after ischemia. Evans Blue dye assay showed that ischemic preconditioning reduced damage to the blood-brain barrier 24 hours after ischemia. This demonstrates the neuroprotective effect of ischemic preconditioning. Western blot assay revealed a significant reduction in protein levels of integrin αβ, vascular endothelial growth factor and its receptor in mice given ischemic preconditioning compared with mice not given ischemic preconditioning 24 hours after ischemia. These findings suggest that the neuroprotective effect of ischemic preconditioning is associated with lower integrin αβand vascular endothelial growth factor levels in the brain following ischemia.

sflk1-IFN-γ双功能蛋白基因重组质粒的构建和表达及生物学活性鉴定

目的:构建表达可溶性血管内皮生长因子受体2(sVEGFR2,在小鼠又称sflk1)与IFN-γ双功能蛋白的重组质粒pcDNA3.1(+)/sflk1-IFN-γ,对表达的sflk1-IFN-γ重组蛋白的生物学活性进行鉴定。方法:以RT-PCR法分别扩增出sflk1与小鼠IFN-γ基因片段,克隆入pcDNA3.1(+)载体,将该重组质粒表达于真核细胞,以ELISA和W estern blotting分别检测胞外及胞内sflk1-IFN-γ双功能重组蛋白的表达,并对该蛋白的生物学活性进行鉴定。结果:构建的pcDNA3.1(+)/sflk1-IFNγ-重组质粒能够在真核细胞中有效表达,且表达的sflk1-IFN-γ重组蛋白兼有sflk1和IFN-γ两者的生物学活性。结论:成功构建和表达了sflk1-IFN-γ双功能蛋白基因重组质粒,为进一步研究该双功能蛋白的抗肿瘤作用打下了基础。

Possible biological and translational significance of mast cells density in colorectal cancer

Mast cells (MCs), located ubiquitously near blood vessels, are descended from CD34+ hematopoietic stem cells. Initially, although their role has been well defined in hypersensitivity reactions, the discovery of their sharing in both innate and adaptive immunity has allowed to redefine their crucial interplay on the regulatory function between inflammatory and tumor cells through the release of mediators granule-associated (mainly tryptase and vascular endothelial growth factor). In particular, in several animal and human malignancies it has been well demonstrated that activated c-Kit receptor (c-KitR) and tryptase (an agonist of the proteinase-activated receptor-2) take pivotal part in tumor angiogenesis after the MCs activation, contributing to tumor cells invasion and metastasis. In this review, we focused on crucial MCs density (MCD) role in colorectal cancer (CRC) development and progression angiogenesis-mediated; then, we will analyze the principal studies that have focused on MCD as possible prognostic factor. Finally, we will consider a possible role of MCD as novel therapeutic target mainly by c-KitR tyrosine kinase inhibitors (imatinib, masitinib) and tryptase inhibitors (gabexate and nafamostat mesylate) with the aim to prevent CRC progression.