Fabrication and Properties of Porous Film / Fabric Composites for Vascular Scaffold Application

A composite vascular scaffold combining textile reinforced structure and biodegradable polymer is introduced, which may possess high porosity and connectivity.Moreover,the porous size could be controlled.The proposed scaffold consists of a warp-knitted poly( ethylene terephthalate)( PET) fabric with well-defined macropores, which is embedded with a porous biodegradable polymer membrane.The aim of this paper is to study the fabrication and properties of porous polymer membrane through optimizing the parameter of composite methods from freeze drying / particle leaching( FD/PL) and gas foaming /particle leaching( GF /PL),subsequently combining with the warp-knitted fabric.Weighing method was utilized to analyze the porosity of the samples and the scanning electron microscope( SEM) images were taken to observe the porous structure of the vascular membrane.In addition,the static contact angle( CA) was measured to estimate the hydrophilicity of the samples, and the tensile testing of the composites was performed on the universal mechanical tester.Furthermore, the water permeability of the membrane was also calculated.The results showed that the porosity and pore connectivity of the vascular membrane were diverse, because of solution concentration, particle size, ratio of content, etc.Meanwhile,the stress-strain curves and the bursting strength showed the different mechanical properties among composite scaffolds in different structures.

Effect of Yiqi Huoxue (益气活血) Herbs on Vascular Endothelial Cells and Platelets in Patients with Chronic Cor Pulmonale

Objective: To observe the clinical effect of Yiqi Huoxue (益气活血, YQHX) herbs in treating the patients with chronic cor pulmonale and to explore its mechanism by determining the relationship of oxida-tion/antioxidation system and how such herbs change on the function of endothelial cells and platelets. Methods: Fifty-eight patients were divided into two groups: conventional therapy group (control group, 28 patients) and convention plus YQHX herbs group (treated group, 30 patients). The control group received conventional management. The treated group were treated with YQHX 150 ml, twice a day, plus the conventional treatment, and the clinical efficacy was recorded. The lipid peroxidation (LPO), superoxide dismutase (SOD), ot-granule membrane protein (GMP140), partial pressure of arterial oxygen (PaO2), partial pressure of carbon dioxide in artery (PaCO2) and circulating endothelial cells (CEC) were measured respectively before and after treatment, and the relationship between various parameters were analyzed. The results were compared with those of 10 healthy subjects got at the same period. Results: (1) The effective rate and PaO2 of the treated group was higher than that of the control group and there were no difference in PaCO2 between the two groups. (2) The levels of LPO, GMP140, CEC in all the patients before therapy were significantly higher than those of the healthy group, and there were marked decrease in the levels of those after treatment (all P<0. 01). On the contrary, the levels of SOD in all the patients before therapy were markedly lower than those in the healthy subjects and increased after treatment, P<0. 01. (3) The increase of SOD in the treated group was significantly more obvious than that of the control group. In the treated group, the decrease of LPO, GMP140, CEC were markedly more obvious than those in the control group (all P<0. 01). (4) The number of CEC, as well as GMP140, was negatively correlated to PaO2(P<0.01) and SOD (P< 0. 01), which was positively correlated to LPO (P<0. 01). There was a positive correlation between CEC and GMP140 (P<0.05). Conclusion: YQHX herbs in treating chronic cor pulmonale proved to be effective by balancing the oxidation and antioxidation, protecting the pulmonary endothelial cells and activated platelets and helpful in treating respiratory failure.

A bio-instructive parylene-based conformal coating suppresses thrombosis and intimal hyperplasia of implantable vascular devices

Implantable vascular devices are widely used in clinical treatments for various vascular diseases. However, current approved clinical implantable vascular devices generally have high failure rates primarily due to their surface lacking inherent functional endothelium. Here, inspired by the pathological mechanisms of vascular device failure and physiological functions of native endothelium, we developed a new generation of bioactive parylene (poly(p-xylylene))-based conformal coating to address these challenges of the vascular devices. This coating used a polyethylene glycol (PEG) linker to introduce an endothelial progenitor cell (EPC) specific binding ligand LXW7 (cGRGDdvc) onto the vascular devices for preventing platelet adhesion and selectively capturing endogenous EPCs. Also, we confirmed the long-term stability and function of this coating in human serum. Using two vascular disease-related large animal models, a porcine carotid artery interposition model and a porcine carotid artery-jugular vein arteriovenous graft model, we demonstrated that this coating enabled rapid generation of self-renewable “living” endothelium on the blood contacting surface of the expanded polytetrafluoroethylene (ePTFE) grafts after implantation. We expect this easy-to-apply conformal coating will present a promising avenue to engineer surface properties of “off-the-shelf” implantable vascular devices for long-lasting performance in the clinical settings.

The role of membrane potential and calcium kinetic changes in the pathogenesis of vascular hyporeactivity during severe shock

OBJECTIVE: To determine the role of membrane potential and intracellular calcium kinetic changes in producing vascular hyporeactivity during severe hemorrhagic shock. METHODS: Rats were subjected to hemorrhagic shock (HS) for 2 hours. The spinotrapezius muscle was prepared for microscopy and the responses of arterioles in the muscle to norepinephrine (NE) were tested. The resting membrane potentials of isolated arterial strips were measured with a microelectrode. Membrane potential and intracellular Ca2+ ([Ca2+]i) changes in isolated arteriolar smooth muscle cells (ASMCs) were determined with fluorescent probes and a confocal microscopy. RESULTS: The arteriolar resting membrane potential was decreased from -36.7 +/- 6.3 mV in control to -29.2 +/- 5.3 mV concurrent with the increase of vasoreactivity to NE at 20 minutes after HS. At 120 minutes post-HS, the resting potential hyperpolarized to -51.9 +/- 9.1 mV, and NE stimulated [Ca2+]i increase was reduced to 50% of the control values during the appearance of arteriolar hyporeactivity, i.e. the NE threshold of the arteriolar response increased 15 fold 2 hours after the onset of hemorrhage as compared with normal animals. The state of vasoreactivity was closely related to the resting potential of vascular smooth muscle in hemorrhagic shock, with a correlation coefficient of 0.96. Treatment with glybenclamide, a selective blocker of ATP-sensitive K+ (KATP) channels, decreased the resting potential, increased NE-stimulated [Ca2+]i increase, and partially restored vasoreactivity in severe hemorrhagic shock. CONCLUSION: The results suggested that membrane hyperpolarization and the reduction of NE-stimulated [Ca2+]i increase in smooth muscle cells appeared to contribute to the vascular hyporeactivity in hemorrhagic shock. The mechanism is likely to involve in KATP channels.

Preparation of Poly(ε-caprolactone)/Poly(ester amide)Electrospun Membranes for Vascular Repair

With adjustable amphiphilicity and anionic/cationic charge,biodegradability and biocompatibility,amino acid-based poly(ester amide)s(PEAs)have drawn attention in the research of tissue engineered vascular grafts.In this work,L-phenylalanine-based PEAs with or without L-lysine were synthesized through polycondensation,and ultrafine fibrous grafts consisted of PEAs and poly(ε-caprolactone)(PCL)in given mass ratios were further prepared via blend electrospinning.Surface characterizations by Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy confirmed the chemical structure,and the wettability indicated that the prepared PCL/PEAs electrospun membranes exhibited less hydrophobic than PCL.Tensile results showed that the PCL/PEAs membranes possessed suitable mechanical properties,which could meet the requirements of artificial blood vessels.Cell culture and hemolytic tests exhibited that the PCL/PEAs electrospun membranes are biocompatible.In general,the electrospun grafts of PCL/PEAs could be applied for vascular repair.

Establishment of the model of vascular endothelial cell membrane chromatography and its preliminary application

A model of vascular endothelial cell membrane chromatography was established by using an ECV304 cell membrane stationary phase (ECV304 CMSP) prepared by immobilizing the ECV304 cell membrane onto the surface of silica carrier. The surface and chromatographic characteristics of ECV304 CMSP were studied. The active component from Caulophyllum robustum was screened by using the model of vascular endothelial cell membrane chromatography. The interaction between the active component and membrane receptor was determined by using a replace experiments. The effect of the active component was tested by using tube formation of ECV304 cell. The results indicated that the model of ECV304 cell membrane chromatograph (ECV304 CMC) can stimulate the interaction between drug and receptor in vitro and the retention characteristics of taspine as active component was similar to that of model molecule in the model of ECV304 CMC. And therefore, taspine acted on VEGFR2 and inhibited the tube formation of ECV304 cell induced by VEGF. This model can be used to screen definite active component as a screening model.

Corneal Neovascularization Suppressed by TIMP2 Released from Human Amniotic Membranes

Purpose: To investigate the effects of culture medium of human amniotic membrane (AM) on corneal neovascularization (CNV) induced by basic fibroblast growth factor (bFGF) in mice.Methods: Culture medium of amniotic membrane was prepared by cultivating AM (with epithelium side up) in EGM basic medium for 3 days, and was collected separately to three groups, e.g. Control (EGM only), AM with epithelium (AM) and AM without epithelium (De-AM). Corneal neovascularization was induced in mice by using micropocket assay with Hydron polymer pellets containing 100 ng bFGF. Migration and proliferation of human umbilical cord vein endothelial cells (HUVEC) were performed in Boyden chambers and by using the CyQUANT fluorescence binding assay respectively.The levels of tissue inhibitors of metalloproteinase 1 and 2 (TIMP1, TIMP2) in culture medium were determined by ELISA assay.Results: CNV induced by bFGF was significantly suppressed by culture medium of amniotic membrane. When the medium was applied as an eyedrop 4 times a day for 7 days,the area of CNV was (2.48±0.76) mm2,(0.64±0.52) mm2 and (1.96±0.65) mm2 incontrol, AM and De-AM group respectively. The migration and proliferation of HUVEC were strongly inhibited by culture medium of AM with epithelium, while the De-AM had no effect on the migration of HUVEC cells. The high level of TIMP2 was found in AM group, but not in De-AM group, while there was no difference in the amount of TIMP1 in medium among three groups.Conclusion: Culture medium of amniotic membrane significantly suppresses the corneal nevovascularization induced by bFGF. The mechanism of which at least in part is that high level of TIMP2 protein secreted or released into the culture medium of AM and inhibition of migration and growth of vascular endothelial cells.

胎衣不下奶牛miRNA-185靶向调控血管内皮生长因子A表达的研究

【目的】通过体内和体外试验检测miRNA-185及血管内皮生长因子A(vascular endothelial growth factor A,VEGFA)在胎衣正常排出和胎衣不下(RFM)奶牛胎盘组织中的差异表达情况及VEGFA在奶牛母体胎盘组织中表达分布,验证及明确miRNA-185和VEGFA与奶牛胎衣不下发生密切相关并存在靶向调节关系,为深入研究miRNA-185在奶牛胎衣不下发生过程中的作用提供理论依据和试验基础。【方法】采用实时荧光定量PCR法检测胎衣正常排出及胎衣不下奶牛母体胎盘组织中miRNA-185及VEGFA基因表达情况,采用荧光原位杂交技术(FISH)检测VEGFA在母体胎盘组织的表达分布及mRNA的差异表达情况;采用双荧光素酶明确miRNA-185与VEGFA的靶向调节关系,采用实时荧光定量PCR及Western blotting检测体外转染miRNA185 mimics、miRNA-185 inhibitor及miRNA-185 NC后VEGFA mRNA及蛋白的差异表达情况。【结果】实时荧光定量PCR结果显示,与胎衣正常排出奶牛相比,胎衣不下奶牛体内miRNA-185及VEGFA的表达水平均极显著下调(P<0.01);荧光原位杂交结果显示,VEGFA mRNA主要在奶牛子宫内膜上皮细胞中表达。VEGFA是miRNA-185的靶向调节蛋白。与阴性对照组相比,转染miRNA-185 mimics后VEGFA的mRNA及蛋白表达水平均极显著下调(P<0.01),转染miRNA-185 inhibitor后VEGFA的mRNA及蛋白表达水平均极显著上调(P<0.01)。【结论】胎衣不下奶牛母体胎盘组织内miRNA-185及VEGFA表达显著降低,VRGFA主要在子宫内膜上皮细胞中表达,miRNA-185可通过靶向调控VEGFA的表达参与奶牛胎衣不下的发生发展。

经皮置管静脉-动脉体外膜氧合支持撤机后血管事件发生情况及相关因素分析

目的分析经皮置管静脉-动脉体外膜氧合支持(V-A ECMO)撤机后的血管事件和发生的临床相关因素。方法回顾性分析郑州大学第二附属医院重症医学科77例成功撤机的V-A ECMO患者。将发生血管事件患者纳入病例组,未发生患者纳入对照组。比较两组患者临床资料,采用多因素分析logistic回归分析V-A ECMO撤机后血管事件的危险因素。结果2015—2023年,共有124例患者成功撤机,其中77例患者被纳入本研究。15例(15/77,19.48%)发生导管相关性深静脉血栓(CaDVT);7例出现动脉血管并发症(9/77,9.09%),其中3例为下肢动脉闭塞,涉及双侧胫前动脉、双侧胫后动脉远端和足背动脉节段,2例为股动脉血栓,均接受了动脉取栓术,2例为股动脉瘤。年龄≥60岁、ECMO前6 h序贯性器官功能衰竭(SOFA)评分、ECMO运行时间、ECMO撤机后感染与血管并发症相关。结论V-A ECMO患者撤机后的血管并发症主要表现为深静脉血栓和动脉并发症。年龄≥60岁、ECMO运行时间和ECMO撤机后感染是撤机后发生血管事件的独立危险因素。

溶酶体相关膜蛋白3通过VEGF/AKT通路抑制PC-3细胞增殖、转移及血管生成

目的探索LAMP3对PC-3细胞增殖、迁移及血管生成的影响。方法Western blot(WB)及RT-PCR检测LAMP3在正常前列腺上皮细胞及前列腺癌骨转移细胞中的表达。构建稳定沉默LAMP3的PC-3细胞,分别使用CCK8、划痕试验、Transwell试验检测LAMP3对PC-3细胞增殖、迁移与侵袭的影响。通过ELISA及血管生成试验检测血管内皮生长因子VEGF、基质金属酶MMP9的表达及HUVEC细胞的血管生成,最后使用WB及RT-PCR检测VEGF、AKT/p-AKT的表达。结果LAMP3在前列腺癌细胞中的表达较正常前列腺上皮细胞明显升高,以PC-3细胞最明显(P<0.05)。沉默LAMP3能抑制PC-3细胞的增殖、迁移与侵袭能力,同时能抑制VEGF、MMP9的表达及PC-3细胞诱导的血管生成,差异有统计学意义(P<0.05)。此外,LAMP3能下调PC-3细胞中VEGF、AKT/p-AKT的表达。结论LAMP3能通过调控VEGF/AKT通路影响PC-3细胞的增殖、转移和血管生成,LAMP3可能是前列腺癌骨转移的潜在治疗靶点之一。

Takeda G蛋白偶联受体5在血管平滑肌细胞增殖和迁移中的作用

目的探讨Takeda G蛋白偶联受体5(Takeda G protein-coupled receptor 5,TGR5)在小鼠血管平滑肌细胞(vascular smooth muscle cells,VSMCs)增殖和迁移中的作用及机制。方法用血小板衍生生长因子(platelet-derived growth factor,PDGF-BB)诱导VSMCs增殖、迁移,以INT-777特异性激活TGR5,CCK-8试剂盒及增殖细胞核抗原(Ki-67)免疫荧光染色用于检测细胞增殖能力,划痕试验用于检测细胞迁移能力。Western blot检测TGR5蛋白水平变化。为探究TGR5在血管内膜增生中的作用,将40只雄性野生型C57BL/6J小鼠随机分为假手术组、内膜损伤组、假手术+UDCA(熊去氧胆酸,TGR5激动剂)组及内膜损伤+UDCA组,每组10只。造模完成后按组分别予以口服普通维持饲料及含0.5%UDCA的普通维持饲料,持续21 d后分别取材,HE染色观察颈动脉内膜增生程度,Ki-67免疫荧光染色观察颈动脉内膜血管平滑肌增殖变化。结果特异性激活TGR5明显降低VSMCs增殖活力及Ki-67阳性细胞率,同时使VSMCs划痕愈合速度减慢。特异性激活TGR5使细胞内UCP2表达增加、活性氧(reactive oxygen species,ROS)水平降低。过氧化氢恢复细胞内ROS水平后,TGR5抑制VSMCs增殖迁移的作用被削弱。激活TGR5能减轻颈动脉损伤后内膜增生。结论TGR5可能通过UCP2改善细胞内氧化应激,从而抑制小鼠VSMCs的增殖和迁移。

探讨体外受精-胚胎移植足月妊娠胎盘屏障的改变

目的使用光镜及透射电镜探究经体外受精-胚胎移植(IVF-ET)助孕足月分娩的胎盘与自然妊娠足月分娩的胎盘的形态学改变。方法选择在郑州大学第三附属医院住院,经IVF-ET助孕分娩的6例胎盘为IVF-ET组,同期住院自然妊娠分娩的10例胎盘为对照组。两组均为足月、初产、无妊娠并发症、择期行剖宫产分娩病例。使用光镜观察两组胎盘中整体结构的改变,并使用透射电镜观察两组胎盘中超微结构的改变。结果IVF-ET组产妇年龄显著高于自然妊娠组[(32.8±1.9)岁vs.(26.7±2.9)岁,P<0.05],两组间孕周、母亲体质量指数(BMI)、新生儿出生体重、胎盘重量和胎盘效率比较均无显著性差异(P>0.05)。光镜下观察,IVF-ET组与自然妊娠组胎盘整体结构未见明显差异。透射电镜观察,与自然妊娠组胎盘相比,IVF-ET组胎盘屏障厚度[(3749.0±1514.0)μm vs.(3379.4±1080.6)μm]、合体滋养细胞基底膜厚度[(66.2±17.5)μm vs.(60.2±15.9)μm]、血管内皮细胞基底膜厚度[(104.2±23.4)μm vs.(103.5±21.9)μm]均呈增加趋势,但差异无统计学意义(P>0.05);IVF-ET组胎盘部分视野中可见到合体滋养细胞顶端微绒毛减少甚至缺失。结论IVF-ET技术并未造成胎盘大体结构上的明显改变,但其胎盘屏障厚度呈增厚趋势,合体滋养细胞顶端微绒毛减少,提示IVF-ET技术可能在一定程度上阻碍了胎儿与母体间的营养物质交换。

REEP5在川崎病中的表达及通过影响线粒体功能对血管内皮细胞早期凋亡的影响

目的探讨REEP5在川崎病患儿中的表达,以及其通过影响线粒体功能对血管内皮早期细胞凋亡的影响。方法选取2020年1月至2023年6月于唐山中心医院就诊的69例川崎病患儿为研究对象,根据是否发生冠状动脉损伤分为冠状动脉损伤组(33例)和非冠状动脉损伤组(36例),选取30例健康体检儿童为健康对照组。酶免疫吸附法检测血清REEP5表达,检测细胞凋亡、血管生成能力、线粒体功能及细胞增殖和蛋白表达。结果与健康对照组相比,REEP5在冠状动脉损伤组及非冠状动脉损伤组中表达显著偏低[(1.05±0.63)比(4.64±1.18)、(3.16±0.85)比(4.64±1.18),P<0.05],且冠状动脉损伤组低于非冠状动脉损伤组(P<0.05)。血小板、红细胞沉降率、白细胞、凝血酶原时间、活化部分凝血活酶时间、纤维蛋白原降解产物、纤维蛋白原与血清REEP5具有显著负相关(P<0.05)。体外细胞实验中:较于对照组,REEP5蛋白和miRNA在模型组中低表达[(1.33±0.15)比(0.34±0.05),(1.24±0.11)比(0.23±0.06),P<0.05],且血管形成数量及血管密度低于对照组(P<0.05)。较于对照组,空白组细胞活力和细胞增殖能力显著降低,LDH活性增加,线粒体膜电位下降,mPTP开放度升高(均P<0.05),细胞早期凋亡增加(P<0.05)。与空白组比较,si-REEP5组细胞活力和细胞增殖能力显著更低,LDH活性增加更显著,线粒体膜电位下降更显著,mPTP开放度显著升高(均P<0.05),细胞早期凋亡显著增加(P<0.05)。与对照组、空白组、si-REEP5组比较,REEP5过表达组细胞活力和细胞增殖能力显著升高,LDH活性显著降低,线粒体膜电位显著升高,mPTP开放度显著下降(均P<0.05),细胞早期凋亡显著降低(P<0.05)。结论在川崎病患儿中,REEP5呈显著低表达,REEP5可通过影响线粒体膜电位和mPTP开放度,影响细胞早期凋亡。

血清sICAM-1、VEGF、sFas水平在急性白血病中的变化及临床价值

目的探讨血清可溶性细胞间黏附分子-1(sICAM-1)、血管内皮生长因子(VEGF)、可溶性跨膜蛋白(sFas)在急性白血病中的变化及临床意义。方法选取2020年5月至2022年5月安岳县人民医院收治的急性白血病患者89例为观察组,另选同时段该院的体检健康者65例为对照组。所有患者入院后给予化疗,对其实施1年电话、门诊随访。比较两组患者血清sICAM-1、VEGF、sFas水平差异,比较不同疗效、不同预后急性白血病患者血清sICAM-1、VEGF、sFas水平差异。绘制受试者工作特征(ROC)曲线,评估相关指标对急性白血病疗效、预后的预测价值。结果与对照组比较,观察组血清sICAM-1、VEGF、sFas水平升高(P<0.05)。与非完全缓解(CR)患者比较,CR患者血清VEGF水平升高(P<0.05)。与预后良好患者比较,预后不良患者血清sICAM-1、VEGF、sFas水平升高(P<0.05)。ROC曲线分析结果显示,VEGF评估急性白血病疗效的曲线下面积(AUC)为0.778,sICAM-1、VEGF、sFas单独与联合评估急性白血病预后的AUC分别为0.793、0.729、0.666、0.889。结论血清sICAM-1、VEGF、sFas在急性白血病患者中水平升高,sICAM-1、VEGF、sFas联合评估急性白血病疗效、预后均有一定临床参考价值。

Challenges and strategies for in situ endothelialization and long-term lumen patency of vascular grafts

Vascular diseases are the most prevalent cause of ischemic necrosis of tissue and organ,which even result in dysfunction and death.Vascular regeneration or artificial vascular graft,as the conventional treatment modality,has received keen attentions.However,small-diameter(diameter<4 mm)vascular grafts have a high risk of thrombosis and intimal hyperplasia(IH),which makes long-term lumen patency challengeable.Endothelial cells(ECs)form the inner endothelium layer,and are crucial for anti-coagulation and thrombogenesis.Thus,promoting in situ endothelialization in vascular graft remodeling takes top priority,which requires recruitment of endothelia progenitor cells(EPCs),migration,adhesion,proliferation and activation of EPCs and ECs.Chemotaxis aimed at ligands on EPC surface can be utilized for EPC homing,while nanofibrous structure,biocompatible surface and cell-capturing molecules on graft surface can be applied for cell adhesion.Moreover,cell orientation can be regulated by topography of scaffold,and cell bioactivity can be modulated by growth factors and therapeutic genes.Additionally,surface modification can also reduce thrombogenesis,and some drug release can inhibit IH.Considering the influence of macrophages on ECs and smooth muscle cells(SMCs),scaffolds loaded with drugs that can promote M2 polarization are alternative strategies.In conclusion,the advanced strategies for enhanced long-term lumen patency of vascular grafts are summarized in this review.Strategies for recruitment of EPCs,adhesion,proliferation and activation of EPCs and ECs,anti-thrombogenesis,anti-IH,and immunomodulation are discussed.Ideal vascular grafts with appropriate surface modification,loading and fabrication strategies are required in further studies.

Complement factor B knockdown by short hairpin RNA inhibits laser-induced choroidal neovascularization in rats

AIM:To evaluate whether recombinant complement factor B(CFB)short hairpin RNA(sh RNA)reduces laserinduced choroidal neovascularization(CNV)in rats.METHODS:Laser-induced rat CNV model was established,and then the animals underwent fundus fluorescence angiography(FFA)and hematoxylin and eosin(HE)staining.On day 3 and 7 after photocoagulation,the expression of CFB and membrane attack complex(MAC)was detected by immunhischemistry.A recombinant CFBsh RNA plasmid was constructed.CFB and scrambled sh RNA plasmids were intravenous injected into rats via the tail vein on the day of laser treatment,respectively.On day 7,the incidence of CNV was determined by FFA,and the expression of CFB and vascular endothelial growth factor(VEGF)in retinal pigment epithelium(RPE)/choroidal tissues was detected by immunhischemistry,Western blot and/or semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR)in CFB and scrambled sh RNA groups.The possible adverse effects of CFB-sh RNA injection were assessed by transmission electron microscopy and electroretinography.RESULTS:FFA and HE results indicated that a laserinduced rat CNV model was successfully established on day 7 after photocoagulation.The expression of CFB and MAC was extremely weak in normal retina and choroid,and increased on day 3 after photocoagulation.However,it started to reduce on day 7.CFB sh RNA plasmid was successfully constructed and induced CFB knockdown in the retinal and choroidal tissues.FFA showed CFB knockdown significantly inhibited incidence of CNV in rats.Moreover,CFB knockdown significantly inhibited the expression of VEGF in RPE/choroidal tissues.CFB sh RNA caused no obvious side effects in eyes.CONCLUSION:CFB knockdown significantly inhibits the formation and development of CNV in vivo through reducing the expression of VEGF,which is a potential therapy target.The alternative pathway of complement activation plays an important role in CNV formation.

Epithelial membrane protein 1 negatively regulates cell growth and metastasis in colorectal carcinoma

AIM:To determine the expression and function of epithelial membrane protein 1(EMP1)in colorectal carcinoma.METHODS:Colorectal samples were taken from cancer lesions and adjacent normal tissue in colorectal cancer patients immediately after endoscopic biopsy.A portion of the sample was either fixed in 4%paraformaldehyde and embedded in paraffin for immunohistochemistry or stored in liquid nitrogen for Western blot.In order to determine protein expression of EMP1in colorectal cancer(n=63)and normal tissue(n=31),semi-quantitative immunohistochemistry and Western blot were utilized.For in vitro studies,the human colorectal cancer cell line SW-480 was maintained in RPMI-1640 medium supplemented with 10%fetal bovine serum.Recombinant lentivirus mediated overexpression of EMP1 in SW-480 cells was quantified by real-time reverse transcription-polymerase chain reaction and Western blot.Control SW-480 cells were transfected with an empty vector.To further study the effect of EMP1 overexpression in SW-480 cells,cell proliferation,apoptosis,migration and invasion assays were conducted.RESULTS:Expression of EMP1 was significantly lower in colorectal cancer tissue than in normal tissue using both immunohistochemistry(39.7%vs 90.3%of tissues,P<0.05)and Western blot(0.126±0.022 vs0.632±0.053,P<0.05).The level of EMP1 protein expression was not correlated with gender,age,or tumor location.Decreased expression of EMP1 was significantly correlated with T stage,lymph node metastasis,clinic stage,and histological grade in patients with colorectal cancer(P<0.05).According to Kaplan-Meier analysis,low EMP1 expression correlated significantly with poor overall five-year survival(34.2%vs 64.0%survival,P<0.05).SW-480 cells transfected with EMP1 had a lower survival fraction,higher cell apoptosis(12.1%±1.3%vs 3.1%±0.6%,P<0.05),a significant decrease in migration and invasion(124.0±17.0 and 87.0±12.0,respectively vs 213.0±29.0 and 178.0±21.0,respectively,P<0.05),higher caspase-9(0.635±0.063 vs0.315±0.032,P<0.05),and lower VEGFC protein expression(0.229±0.021 vs 0.519±0.055,P<0.05)relative to cells not transfected with EMP1.CONCLUSION:Low EMP1 expression in colorectal cancer is associated with increased disease severity,suggesting that EMP1 may be a negative regulator of colorectal cancer.

Iododeoxyuridine uptake in proliferating smooth musc le cells:an in vitro model to assess drug effects on intimal hyperplasia

Purpose To assess the maximum uptake of Iododeo xyur idine (IUdR) by proliferating smooth muscle cells in vitro to determine the opti mal concentration to be administrated in an in vivo experiment. The long-term g oal is to utilize radioactive IUdR to inhibit smooth muscle cell proliferation a nd restenosis of arteries after balloon angioplasty in vivo. Methods Porcine smooth muscle cells (SMCs) were cultured in 5% FBS medium and stim ulated to proliferate by the addition of medium containing 10% FBS and insulin. IUdR was added at 5 μM, 10 μM, 20 μM, 30 μM, 40 μM, respectively, in prolif erating SMCs with control for 1, 3, 5, 7 day incubation. Fluorescence Activated Cell Scanning (FACS) was performed after the SMCs were harvested and double-sta ined with FITC-conjugated anti-IUdR antibody (B44) and propidium iodide (PI). The ratio of IUdR-labeled cells to total cell population for each IUdR concentr ation and duration was determined by FACS. All data were repeated three times at each time point. The doubling times, growth curve and cell density of the proli ferating SMCs were investigated using Beckman Coulter Particle Counter and digit al microscopy. Results The percentage of proliferating SMCs uptaking IUdR incr eased from 1 to 5 days incubation with all concentrations of IUdR; In day 5, the uptake rate reached the peak value, then decreased by 7 days. IUdR uptake on d ay 5 was higher with concentrations of 10 μM and 20 μM. The doubling times of the SMCs were prolonged with IUdR concentration increasing, while the proliferat ing cell number and density compared with control decreased obviously by day 5 ( P<0.05).Conclusion The peak time to uptake IUdR was 5 days and optimal concentration of IUdR was between10 μM to 20 μM for proliferating SMCs to upta ke in vitro. IUdR itself could inhibit the SMCs’ proliferation and the inhibito ry effect was related to the concentration.[

改良内瘘穿刺法在疑难动静脉内瘘患者中的效果评价

目的 探讨改良内瘘穿刺法在疑难动静脉内瘘患者中的应用效果。方法 将2020年6月—2022年6月在江苏省人民医院进行血液透析的80例疑难自体内瘘患者随机分为对照组和试验组,每组40例。对照组采用常规穿刺法,试验组采用改良内瘘穿刺法。比较2组患者穿刺效果、并发症发生率及满意度。结果 2组穿刺10、40次后的内瘘血管内膜壁厚度均较同组穿刺前增加,且对照组穿刺40次后的内瘘血管内膜壁厚度大于试验组穿刺40次后,差异均有统计学意义(P<0.05)。试验组患者一次性穿刺成功率为99.21%,高于对照组的95.62%,差异有统计学意义(P<0.05);试验组患者疼痛评分为(1.60±0.52)分,低于对照组的(2.40±0.97)分,差异有统计学意义(P<0.05)。试验组患者血肿发生率为0.04%,低于对照组的1.90%,差异有统计学意义(P<0.05);试验组穿刺点渗血发生率为0.93%,与对照组的1.46%比较,差异无统计学意义(P>0.05)。试验组患者并发症总发生率低于对照组,差异有统计学意义(P<0.05);试验组患者满意度评分高于对照组,差异有统计学意义(P<0.05)。结论 改良内瘘穿刺法能够减轻疑难动静脉内瘘患者的血管穿刺痛苦,降低血肿或渗血发生率,减少并发症的发生,对内瘘血管有保护作用。

不引起管腔狭窄的血管中膜钙化

血管钙化与心血管事件发生密切相关,主要包括内膜钙化和中膜钙化2种形式。由于中膜钙化不引起管腔狭窄,早期认为其没有临床意义,对其研究较少。近年来研究表明中膜钙化与心血管疾病死亡率密切相关。因此,临床中应注意发现和识别病变中的中膜钙化,明确其对局部和系统血管弹性的影响及与糖尿病神经病变的联系,关注其在心血管疾病中的作用。总结中膜钙化的发生机制、病变特点、诊断方法、致病机制、血流动力学变化及与内膜钙化的区别和联系具有重要的临床意义。