2型糖尿病患者单核细胞-内皮粘附功能的改变(摘要)

目的 :观察 2型糖尿病人外周血单个核细胞对培养的血管内皮细胞的粘附性。方法 :采用单核细胞 -内皮细胞粘附试验检测 5 2例 2型糖尿病患者、3 2例健康志愿者的单核细胞 -血管内皮粘附功能。结果 :(1)糖尿病组外周血单个核细胞 -内皮细胞粘附率显著高于正常对照组 (P <0 .0 0 1) ,(2 )经多元回归分析 ,空腹血糖与单核细胞粘附率存在独立的相关关系 (P <0 .0 0 1)。结论 :2型糖尿病人外周血单个核细胞对培养的血管内皮细胞粘附性增强 ,高血糖等代谢异常与此粘附性增强有关。

结肠腺瘤息肉蛋白突变经PI3K/AKT信号通路影响犬肾细胞MDCK的细胞粘附(英文)

结肠腺瘤息肉蛋白(APC)是一个肿瘤抑制因子,它不仅参与Wnt信号通路的传导,而且对细胞粘附、细胞骨架的组织和迁移等都有影响.APC突变发生于大多数结肠癌中.为了探讨APC突变对细胞粘附的影响及机制,本研究利用细胞粘附实验分析了MDCK-APC-N1和对照MDCK-GFP稳定表达细胞株系的细胞粘附情况.实验结果显示,在MDCK细胞中过表达APC-N1导致细胞-细胞间的粘附减少,细胞-基质间的粘附增加.荧光定量PCR和Western印迹实验表明,在MDCKAPC-N1细胞中,E-cadherin表达水平降低,CD29、P-FAK(Y397)、β-catenin和P-AKT(T308)表达水平升高.在MDCK-APC-N1细胞中,敲减β-catenin导致E-cadherin表达量升高,而CD29表达没有明显变化.进一步利用PI3K抑制剂LY294002处理MDCK-APC-N1细胞,结果发现,E-cadherin表达量明显升高,CD29表达量明显降低.这些结果揭示,APC-N1可活化PI3K/AKT信号通路,进而改变粘附蛋白E-cadherin和CD29影响细胞粘附.

乳化异氟醚对人体外循环血清培养人血管内皮细胞ICAM-1表达变化的影响

目的探讨人体外循环(CPB)血清诱致培养人血管内皮细胞ICAM-1表达及乳化异氟醚对其的影响。方法采用CPB血清致伤培养血管内皮细胞模型,以免疫组化方法观察培养血管内皮细胞ICAM-1表达量。结果各组血管内皮细胞ICAM-1的表达随着时间的延长均呈进行性升高。各时段小牛血清(FBS)组血管内皮细胞ICAM-1表达量最低(P<0.01)。与CPB前血清组相比,CPB后血清组及乳化异氟醚组1h、2h、4h、8h、12h时的ICAM-1表达水平显著升高(P<0.01)。乳化异氟醚组在1h、2h、4h、8h升高的幅度与CPB后血清组相比明显减少(P<0.01),在12h时的ICAM-1表达水平与CPB后血清组相比无显著性差异(P>0.05)。结论CPB显著增加ICAM-1的表达,乳化异氟醚在一定时间内可显著减轻CPB对内皮细胞的影响,减少ICAM-1的表达,对于体外循环后全身炎症反应可能有保护作用。

山楂对人血管内皮细胞的作用

目的 : 探讨山楂对人血管内皮细胞的保护作用及其机制。方法 : 通过原代培养人脐静脉内皮细胞 (EC) ,从细胞形态学、细胞生长状态、细胞内乳酸脱氢酶 (LDH)释放、内皮细胞对单核细胞 (MC- EC)的粘附作用、以及低密度脂蛋白脂质过氧化 (TBARS)值测定方面 ,观察不同剂量山楂对氧化修饰的低密度脂蛋白 (Ox LDL)引起的 EC损伤的影响和对 EC的直接作用。结果 : Ox LDL对 EC具有明显的损伤作用 ,Ox LDL组的细胞存活率低于正常对照组 (P<0 .0 5) ,细胞内 LDH释放率和 MC- EC粘附率均高于正常对照组 (P<0 .0 5) ;山楂可以有效地抑制 Ox LDL对 EC的损伤作用 ,三个剂量的山楂 +Ox LDL各组的细胞存活率均显著高于 Ox LDL 组 (P<0 .0 5) ,LDH释放率、MC- EC粘附率及 TBARS值均显著低于 Ox LDL组 (P<0 .0 5) ,呈剂量效应关系。单纯山楂组的中、高剂量组 MC- EC粘附率均显著低于正常对照组 (P<0 .0 5)。结论 : 山楂可以有效地保护人内皮细胞免受 Ox LDL的损伤 ,其机制与山楂的抗氧化作用和对

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.

Synthesis and anti-shock activity of divalent β-D-galactopyranosyl-(1→4)-β-D-glucopyranosides

Three divalent β-D-galactopyranosyl-（1 → 4）-β-D-glucopyranosides were synthesized using acetylated lactosyl bromide as donor and polyethylene glycols with different polymeric degree （n = 4, 5, 6） as linker, and evaluated for in vivo inhibitory activity to leukocyte-endothelial cell adhesion on severe bum-shock rats. The result showed that the length of linkers had apparent influence on anti cell adhesion activity.

Suppression of matrix metalloproteinase-2 via RNA interference inhibits pancreatic carcinoma cell invasiveness and adhesion

AIM:To investigate the inhibitory effects of RNA interference (RNAi) on expression of matrix metalloproteinase-2 (MMP-2) gene and invasiveness and adhesion of human pancreatic cancer cell line,BxPC-3.METHODS:RNAi was performed using the vector (pGPU6)-based small interference RNA (siRNA) plasmid gene silence system to specifically knock down MMP-2 expression in pancreatic cancer cell line,BxPC-3. Four groups of different specific target sequence in coding region of MMP-2 and one non-specific sequence were chosen to construct four experimental siRNA plasmids of pGPU6-1,pGPU6-2,pGPU6-3 and pGPU6-4,and one negative control siRNA plasmid of pGPU6 (-). MMP-2 expression was measured by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. Cell proliferation and apoptosis were examined by methyl thiazolyl tetrazolium (MTT) and flow cytometry,respectively. The abilities of adhesion and invasion were detected by cell adhesion assay and cell invasion assay using Transwell chambers.RESULTS:The expression of MMP-2 was inhibited and the inhibitory effects of different sequence varied. pGPU6-1 group had the most efficient inhibitory effect,followed by pGPU6-2 and pGPU6-3 groups.Invasiveness and adhesion were more significantly reduced in pGPU6-1,pGPU6-2 and pGPU6-3 groups as compared with pGPU6 (-) and blank control groups. However,no difference concerning cell proliferation and apoptosis was observed after transfection between experiment groups and control groups.CONCLUSION:RNAi against MMP-2 successfully inhibited the mRNA and protein expression of MMP-2 in the pancreatic cancer cell line,BxPC-3,leading to a potent suppression of tumor cell adhesion and invasion without affecting cell proliferation and apoptosis. These findings suggest that the RNAi approach towards MMP-2 may be an effective therapeutic strategy for the clinical management of pancreatic tumor.

急性脑梗死患者血清sELAM-1、IL-1β、TNF-α水平测定

目的:探讨急性脑梗死与血清炎性因子sELAM- 1、IL -1β、TNF -α之间的关系。方法:采用ELISA双抗体夹心法对 40例急性脑梗死患者发病后第 1d、第 5d、第 14d和 40例正常人的血清sELAM、IL 1β、TNF α水平进行测定并对比分析。结果:急性脑梗死患者血清sELAM、IL 1β、TNF α水平在发病第 1d、第 5d、第 14d较对照组升高,并随病程的延长呈下降趋势,但仍未达正常水平 (P均 <0. 01)。且sELAM 1与IL -1β、IL- 1β与TNF- α、TNF -α与sELAM -1间均呈正相关关系,r分别为 0. 801、0. 818及 0. 820(P均<0. 01)。结论:血清sELAM 1、IL- 1β和TNF- α参与了急性脑梗死的病理生理过程和缺血性脑损害。

动脉粥样硬化斑块的研究进展

本文阐述了动脉粥样硬化斑块的形成和发展以及内皮细胞

丹红注射液联合体外反搏对急性冠脉综合征患者PCI术后炎症因子的影响

目的探讨丹红注射液联合体外反搏治疗对急性冠脉综合征患者经皮冠状动脉介入术(PCI)后血浆炎症因子水平的影响。方法选取2012年11月~2014年8月在青岛大学附属医院因急性冠脉综合征行PCI手术,后转入青岛大学附属心血管病医院治疗的患者118例,其中男性68例,女性50例,按随机数表法分为Ⅰ组(n=38)、Ⅱ组(n=40)、Ⅲ组(n=40)。三组患者均给予常规治疗,Ⅰ组给予常规治疗,Ⅱ组在常规治疗基础上给予0.9%氯化钠250 ml或5%葡萄糖250 ml+丹红注射液40 ml静脉滴注1/日,Ⅲ组在常规治疗基础上给予0.9%氯化钠250 ml或5%葡萄糖250 ml+丹红注射液40 ml静脉滴注1/日及体外反搏治疗(1h/次,1/日),连续治疗14d。PCI术前1d、PCI术后1天(24h±2h)、治疗14d时测定高敏C反应蛋白(hs CRP)、血清细胞间粘附分子-1(ICAM-1)和血管细胞粘附分子-1(VCAM-1)。结果Ⅰ组有2例、Ⅱ组有1例、Ⅲ组有3例由于不能耐受体外反搏或病情加重中途退出试验。与PCI术前1d比较,PCI术后1d三组hs CRP、ICAM-1和VCAM-1均升高,差别有统计学意义(P<0.05);用药14d时,与Ⅰ组相比,Ⅱ组、Ⅲ组hs CRP、ICAM-1、VCAM-1较前降低,差异有统计学意义(P<0.05);与Ⅱ组相比,Ⅲ组hs CRP、ICAM-1、VCAM-1降低,差别有统计学意义(P<0.05)。结论丹红联合体外反博能降低急性冠脉综合征患者PCI术后的炎症因子水平。

BCL2 inhibits cell adhesion, spreading, and motility by enhancing actin polymerization

BCL2 is best known as a multifnnctional anti-apoptotic protein. However, little is known about its role in cell- adhesive and motility events. Here, we show that BCL2 may play a role in the regulation of cell adhesion, spreading, and motility. When BCL2 was overexpressed in cultured murine and human cell lines, cell spreading, adhesion, and motility were impaired. Consistent with these results, the loss of Bcl2 resulted in higher motility observed in Bcl2- null mouse embryonic fibroblast （MEF） cells compared to wild type. The mechanism of BCL2 regulation of cell adhesion and motility may involve formation of a complex containing BCL2, actin, and gelsolin, which appears to functionally decrease the severing activity of gelsolin. We have observed that the lysate from MCF-7 and NIH3T3 cells that overexpressed BCL2 enhanced actin polymerization in cell-free in vitro assays. Confocal immunofluorescent localization of BCL2 and F-actin during spreading consistently showed that increased expression of BCL2 resulted in increased F-actin polymerization. Thus, the formation of BCL2 and gelsolin complexes （which possibly contain other proteins） appears to play a critical role in the regulation of cell adhesion and migration. Given the established correlation of cell motility with cancer metastasis, this result may explain why the expression Of BCL2 in some tumor cell types reduces the potential for metastasis and is associated with improved patient prognosis.

Systemic inflammatory response following acute myocardial infarction

Acute cardiomyocyte necrosis in the infarcted heart generates damage-associated molecular patterns, activating complement and toll-like receptor/interleukin-1 signaling, and triggering an intense inflammatory response. Iuflammasomes also recognize danger signals and mediate sterile inflammatory response following acute myocardial infarction （AMI）, Inflammatory response serves to repair the heart, but excessive inflammation leads to adverse left ventricular remodeling and heart failure. In addition to local inflammation, profound systemic inflammation response has been documented in patients with AMI, which includes elevation of circulating inflammatory cytokines, chemokines and cell adhesion molecules, and activation of peripheral leukocytes and platelets. The excessive inflammatory response could be caused by a deregulated immune system. AMI is also associated with bone marrow activation and spleen monocytopoiesis, which sustains a continuous supply of monocytes at the site of inflammation. Accumulating evidence has shown that systemic inflammation aggravates atherosclerosis and markers for systemic inflammation are predictors of adverse clinical outcomes （such as death, recurrent myocardial in- farction, and heart failure） in patients with AMI.

Department of Etiology and Department of Pharmacology

In order to understand the role of transmembrane signal transduction of host cells in the early steps of infection,the adherence of E. coli to HEp 2 cells and the change of activity of phospholipase C γ (PLC γ) induced by the adherence were investigated.The adherence of enteropathogenic E.coli (EPEC), strain E.7, induced a significant increase of inositol triphosphat (IP 3) level in HEp 2 cells. The adherence of the bacteria and the increase of IP 3 was kinetically correlated. Whereas the increase of IP 3 level induced by the adherence of the control strain EPEC (H511), a non piliated strain, was much meager than that by E7, a piliated strain. The results highlighted an important role of transmembrane signals like IP 3 in the pathogenesis of EPEC.

Characterization of hepatic progenitors from human fetal liver during second trimester

AIM： To enrich hepatic progenitors using epithelial cell adhesion molecule （EpCAM） as a marker from human fetal liver and investigate the expression of human leukocyte antigen （HLA） and their markers associated with hepatic progenitor cells. METHODS： EpCAM ＋ve cells were isolated using magnetic cell sorting （MACS） from human fetuses （n = 10） at 15-25 wk gestation. Expression of markers for hepatic progenitors such as albumin, alpha-fetoprotein （AFP）, CD29 （integrin ~1）, CD49f （integrin c^6） and CD90 （Thy 1） was studied by using flow cytometry, immunocytochemistry and RT-PCR; HLA class Ⅰ （A, B, C） and class Ⅱ （DR） expression was studied by flow cytometry only. RESULTS： FACS analysis indicated that EpCAM ＋ve cells were positive for CD29, CD49f, CD90, CD34, HLA class I, albumin and AFP but negative for HLA class Ⅱ （DR） and CD45. RT PCR showed that EpCAM ＋ve cells expressed liver epithelial markers （CK18）, biliary specific marker （CK19） and hepatic markers （albumin, AFP）. On immunocytochemical staining, EpCAM ＋ve cells were shown positive signals for CK18 and albumin. CONCLUSION： Our study suggests that these EpCAM ＋ve cells can be used as hepatic progenitors for cell transplantation with a minimum risk of alloreactivity and these cells may serve as a potential source for enrichment of hepatic progenitor.

αV integrin:A new gastrin target in human pancreatic cancer cells

AIM： To analyse αv integrin expression induced by gas- trin in pancreatic cancer models.METHODS： αv integrin mRNA expression in human pan- creatic cancer cells was analysed using a ＂cancer genes＂ array and confirmed by real-time reverse transcription- polymerase chain reaction （PCR）. Western blotting and semi-quantitative immunohistochemistry were used to examine protein levels in human pancreatic cancer cell lines and pancreatic tissues, respectively, The role of αv integrin on gastrin-induced cell adhesion was examined using blocking anti-αv integrin monoclonal antibodies. Adherent cells were quantified by staining with crystal violet.RESULTS： Using a ＂cancer genes＂ array we identi- fied c^v integrin as a new gastrin target gene in human pancreatic cancer cells. A quantitative real-time PCR approach was used to confirm αv integrin gene expression. We also demonstrate that Src family kinases and the PI 3-kinase, two signalling pathways specifically activated by the CCK-2 receptor （CCK2R）, are involved in gastrin-mediated αv integrin expression. In contrast, inhibition of the ERK pathway was without any effect on αv integrin expression induced by gastrin. Our results also show that gastrin modulates cell adhesion via αv integrins. Indeed, in vitro adhesion assays performed on fibronectin show that gastrin significantly increases adhesion of pancreatic cancer cells. The use of blocking anti-αv integrin monoclonal antibodies completely reversed the increase in cell-substrate adhesion induced by gastrin. In addition, we showed in vivo that the targeted CCK2R expression in the pancreas of Elas-CCK2 mice, leads to the overexpression of αv integrin. This process may contribute to pancreatic turnout development observed in these transgenic animals.CONCLUSION： αv integrin is a new gastrin target in pancreatic cancer models and contributes to gastrin effects on cell adhesion.

Neurogenesis by Activation of Inherent Neural Stem Cells in the Rat Hippocampus after Cerebral Infarction

Objective To investigate the changes of neural stem cells (NSCs) in the rat hippocampus after cerebral infarction (CI) and to evaluate the neurogenesis caused by the activation of NSCs. Methods CI models of rats were made and rats were assigned to 6 groups: sham-operated, 1 day, 3 days, 7 days, 14 days, and 28 days after CI. The dynamic expression of bromodeoxyuridine (BrdU), polysialylated neural cell adhesion molecule (PSA-NCAM), glial fibrillary acidic protein (GFAP), and neuronal nuclear antigen (NeuN) were determined by immunohistochemistry and immunofluorescence staining. BrdU was used to mark the proliferated NSCs. PSA-NCAM was used to mark the plasticity of activated NSCs. GFAP and NeuN were used to mark the differentiated NSCs. Results Compared with the controls, the number of BrdU+ cells in the hippocampus increased significantly at 1 day after CI (P<0.05), reached peak at 7 days after CI (P<0.05), decreased but still elevated compared with the controls at 14 days after CI (P<0.05), and nearly unchanged at 28 days after CI. The number of BrdU+/PSA-NCAM+ cells increased significantly at 7 days after CI (P<0.05), reached peak at 14 days after CI (P<0.05), and decreased but still elevated compared with the controls at 28 days after CI (P<0.05). The number of BrdU+/PSA-NCAM+ cells was equal to 60% of the number of BrdU+ cells in all the same period. The number of BrdU+/NeuN+ cells in the hippocampus increased significantly at 14 days after CI (P<0.05) and reached peak at 28 day after CI (P<0.05). The number of BrdU+/GFAP+cells in the hippocampus nearly unchanged after CI. Conclusion CI can stimulate the proliferation of inherent NSCs, and most proliferated NSCs may differentiate into neurons and represent neural plasticity.

Role of integrin β1 in sensitivity to chemotherapy of pulmonary adenocarcinoma A549

Objective:The aim of our study was to investigate the effect of integrin β1 on influencing the sensitivity to chemotherapy of human pulmonary adenocarcinoma A549 cells.Methods:Human pulmonary adenocarcinoma A549 multicellular spheroids(MCS) were constructed with three dimensional cell culture methods.Cell counting using blood cell counter was employed to detect the sensitivity to ADM of A549 MCS before and after blocking integrin β1;integrin β1 expression of A549 MCS and cell apoptosis was detected by flow cytometry.Results:A549 MCS were successfully established.The integrin β1 expression of A549 MCS elevated with the concentration of ADM(< 0.02 μg/mL).Blocking of integrin β1 lead to higher sensitivity to ADM,and IC50 decreased from 0.19 μg/mL to 0.11 μg/mL,and apoptosis rate increased from(15.81 ± 1.87)% to(30.14 ± 2.89)%.Conclusion:The cell adhesion molecules integrin β1 could influence the sensitivity to chemotherapy of A549 MCS and inhibiting of cell apoptosis might be its mechanism.

Morphological Evaluation of Adhesion and Proliferation of Osteoblast Like Cells Grown on Gelatin/Genipin Scaffold

This work focusing on studying the biocompatibility and the effect of gelatin porous scaffold on the characteristics of human osteoblast like cells, including proliferation, adhesion, scaffold-cell interaction and its potential to induce bone regeneration. Osteoblast like cells were seeded on gelatin/genipin scaffolds for 7, 14 and 21 days. Cell proliferation assay, light microscopy, transmission electron microscopy and high resolution scanning electron microscopy were carried to evaluate cell viability, cell adhesion and the production of extracellular matrix. Cell proliferation assay showed a high biocompatibility of the material. High resolution scanning electron microscopy and light microscopy showed a strong adhesion of MG63 ceils on the surface of gelatin scaffold and high penetration in the macroporosities of the material. TEM analysis showed an intense production of extracellular matrix protein. In vitro analysis indicated a good biocompatibility of the scaffold and presents it as a potential candidate material for tissue engineering.