经冠状动脉移植自体骨髓单个核细胞和间充质干细胞对急性心肌梗死模型心功能的改善

目的:为保护濒死心肌提供机会窗口,对比观察经冠脉移植自体骨髓单个核细胞或间充质干细胞后,实验性急性心肌梗死动物心功能变化及心肌组织核转录因子кB、心肌细胞凋亡情况。方法:实验于2005-03/2006-11在河北省人民医院实验中心完成。选用24只雄性冀中白猪,随机数字表法分为4组:正常对照组、模型组、单个核细胞组、间充质干细胞组,6只/组。①24只猪均以盐酸氯胺酮200mg臀部肌肉注射麻醉后,分别于各自右侧股骨抽取骨髓20mL,采用Fercoll法分离获得骨髓单个核细胞,加入胶体金溶液,培养12～16h待用。分离过程中取出含有骨髓单个核细胞成分的细胞层,常规培养传代,每3d换液1次,贴壁生长细胞即为骨髓间充质干细胞,加入胶体金溶液,培养24h待用。②除正常对照组外,其余各组均经导管球囊封闭第一对角支以远的前降支,复制猪急性心肌梗死模型。单个核细胞组、间充质干细胞组均于造模后立即开通前降支,分别经球囊注入预先分离的骨髓单个核细胞6×108个、间充质干细胞6×108个。模型组造模后于梗死1h开通前降支,经球囊注入磷酸盐缓冲液10mL。③各组分别于术前及术后4周经心脏超声检测心功能,取材行病理学检查、心肌组织核转录因子кB的免疫组织化学检测及心肌细胞凋亡检测。结果:24只雄性白猪均进入结果分析。①心功能变化:术前各组左心室收缩末内径、左心室舒张末内径、左心室射血分数、短轴缩短率基本相似。移植术后4周,正常对照组、单个核细胞组、间充质干细胞组左心室舒张末内径均明显低于模型组(F=4.68,P=0.01),左心室射血分数及短轴缩短率均明显高于模型组(F=5.14,P=0.01;F=3.32,P=0.04),各组左心室收缩末内径差异无显著性意义(F=1.64,P=0.21)。②心肌组织病理学改变:电镜下单个核细胞组、间充质干细胞组在梗死边缘区可见有胶体金颗粒的不成熟的心肌细胞,胞质中散在肌丝结构,肌丝排列紊乱不规则。③心肌组织核转录因子кB阳性率表达:与模型组比较,单个核细胞组、间充质干细胞组的梗死边缘区核转录因子кB阳性率明显降低(F=25.59,P=0.0001);正常心肌区核转录因子кB阳性率亦明显降低(F=18.20,P=0.0001)。④心肌细胞凋亡检测结果:与模型组比较,单个核细胞组、间充质干细胞组在心肌梗死区细胞凋亡率均明显降低(F=6.63,P=0.0027),梗死边缘区细胞凋亡率亦明显降低(F=36.07,P=0.0001)。正常心肌区单个核细胞组细胞凋亡率与模型组基本相似(F=9.69,P=0.004),但间充质干细胞组有所降低。⑤心功能与心肌细胞凋亡及心肌组织NF-кB的相关性:急性心肌梗死4周时,左心室射血分数与心肌细胞凋亡、心肌组织核转录因子кB均呈负相关(r=0.613,P=0.001;r=-0.437,P=0.033)。心肌细胞凋亡与心肌组织核转录因子кB呈正相关(r=0.672,P=0.002)。结论:经冠脉移植骨髓单个核细胞和间充质干细胞均可改善实验性急性心肌梗死动物的心功能,与梗死边缘区核转录因子кB表达降低及心肌细胞凋亡减少有关。骨髓单个核细胞移植的促血管增生作用优于间充质干细胞移植。     

Brain diffusion tensor MRI in systematic lupus erythematosus: A systematic review

Diffusion tensor imaging (DTI) maps the brain's microstructure by measuring fractional anisotropy (FA) and mean diffusivity (MD). This systematic review describes brain diffusion tensor Magnetic resonance imaging (MRI) studies in systemic lupus erythematosus (SLE).The literature was reviewed following the PRISMA guidelines and using the terms “lupus”, “systemic lupus erythematosus”, “SLE”, “diffusion tensor imaging”, “DTI”, “white matter” (WM), “microstructural damage”, “tractography”, and “fractional anisotropy”; the search included articles published in English from January 2007 to April 2017. The subjects included in the study were selected according to the ACR criteria and included 195 SLE patients with neuropsychiatric manifestation (NPSLE), 299 without neuropsychiatric manifestation (non-NPSLE), and 423 healthy controls (HC). Most studies identified significantly reduced FA and increased MD values in several WM regions of both NPSLE and non-NPSLE patients compared to HC. Subclinical microstructural changes were observed in either regional areas or the entire brain in both the non-NPSLE and NPSLE groups.     

骨髓间充质干细胞向胰岛素分泌细胞诱导分化的动态观察

目的:探讨体外诱导骨髓间充质干细胞向胰岛素分泌细胞分化的可能性,并观察其动态变化。方法:实验于2005-09/2007-03在山东大学齐鲁医院完成。①标本来源:骨髓标本15例来自山东大学齐鲁医院成人骨髓检查结果正常者,均签署捐献同意书。②实验方法:无菌条件下取骨髓2.0～5.0mL,采用percoll分离液和贴壁法获得纯化的成人骨髓间充质干细胞。③实验评估:流式细胞仪行细胞表面抗原检测,在适当的条件下诱导其分化为成骨细胞和脂肪细胞。采用两步法向胰岛素分泌细胞诱导,观察其在碱性成纤维细胞生长因子、活化素A、胰岛素样生长因子、尼克酰胺等因子刺激下向胰岛素分泌细胞分化的动态变化。双硫踪染色鉴定胰岛样细胞团,酶联免疫吸附试验检测细胞分泌胰岛素的情况,RT-PCR检测胰岛细胞特异基因的表达。结果:①骨髓间充质干细胞的生长特性及免疫表型:分离培养获得的贴壁细胞,呈形态均一的梭形,流式细胞仪检测CD34、CD45表达阴性,CD29、CD44表达阳性。②向成骨细胞和脂肪细胞的诱导分化:此类细胞经茜素红染色、油红O染色均呈阳性,可诱导分化为成骨细胞和脂肪细胞。③向胰岛素分泌细胞的诱导分化:第1步诱导后出现细胞簇,双硫腙染色不着色,胰岛素分泌量少,仅检测到PDX-1基因的表达,证实其为胰岛前体细胞。第2步诱导后细胞簇数目逐渐上升,至诱导14d大部分细胞簇经双硫腙染色都呈红色。④诱导后培养上清中胰岛素含量:诱导第3,7,14,21天的胰岛素分泌量分别为(15.3±4.9),(34.1±5.6),(40.4±5.3),(39.8±5.1)mU/L。⑤胰岛细胞特异基因的表达:诱导7d仅检测到PDX-1基因的表达,insulin1、insulin2和Glut2基因均不表达。诱导14,21d检测到insulin2、PDX-1基因表达,insulin1基因弱表达,Glut2基因不表达。结论:体外分离、纯化得到的骨髓间充质干细胞诱导7d可分化出胰岛前体细胞,不具功能性;诱导14d后可成功地分化出成熟的具有功能性的胰岛素分泌细胞。     

Adipocyte insulin receptor binding and lipogenesis at term in normal pregnancy

Lipogenesis in subcutaneous abdominal adipocytes during late human pregnancy was investigated by studying insulin receptor binding, 3-O-methyl-(14C-(U]-glucose flux and incorporation of (14C(U]-glucose into CO2 (oxidation) and total lipids (lipogenesis) in adipocytes from 18 health pregnant women undergoing Caesarean section at term, and 19 non-pregnant women undergoing gynaecological surgery. The cell diameter and fasting insulin were increased in the pregnant women, compared with controls (P less than 0.01 and P less than 0.05, respectively). The insulin receptor binding, 3-O-methyl-glucose flux, and basal oxidation were similar in both groups. Basal lipogenesis was higher in adipocytes from pregnant women than from controls (P less than 0.05), but the maximally stimulated increment was similar in both groups. Basal and maximally stimulated lipogenesis correlated positively with the cell diameter (P less than 0.001 and P less than 0.05, respectively). The findings indicate that lipogenesis in subcutaneous abdominal adipocytes from pregnant women is increased due to post-receptor events and that adipocytes do not contribute to the insulin resistance in late pregnancy.     

代谢综合征的临床相关指标

目的:分析超氧化物歧化酶、丙二醛、瘦素、脂联素等指标在代谢综合征发病过程中的作用。方法:应用计算机检索中国医院知识仓库(CHKD)数据库1998-01/2005-12相关代谢综合征方面的文献,检索词"代谢综合征",限定文献语言种类为中文。对资料进行初审,选取包括超氧化物歧化酶、丙二醛、瘦素、脂联素的文献,开始查找全文。纳入标准:临床对照研究。排除标准:综述性研究,重复性研究。共检索到24篇关于代谢综合征与超氧化物歧化酶、丙二醛、瘦素、脂联素等指标的文献,最终纳入16篇符合标准的文献。结果:研究发现超氧化物歧化酶、丙二醛、瘦素、脂联素等指标与代谢综合征的关系密切。丙二醛的高低可间接反映机体细胞受氧自由基损伤的程度,超氧化物歧化酶的检测可间接反映机体抗氧化的能力,代谢综合征患者超氧化物歧化酶活性降低、丙二醛堆积,提示其抗氧化能力减弱。代谢综合征患者多存在高瘦素、低脂联素血症,提示脂联素、瘦素在代谢综合征发生发展过程中发挥了重要作用,低脂联素血症、高瘦素血症可能是代谢综合征的重要组成成分。结论:动态观察血清瘦素、脂联素、超氧化物歧化酶水平可以了解代谢综合征发生发展的过程,亦可以作为疗效观测的指标。     

The effect of gastrin on basal and aminoacid-stimulated insulin and glucagon secretion in man

Abstract. The effect of gastrin on basal and aminoacid-stimulated glucagon and insulin secretion was studied in eleven normal young subjects. The concentrations of glucagon, insulin and gastrin in plasma or serum were measured radioimmunochemically. The results of amino-acid-stimulation were compared to those obtained during a protein-rich meal.
Intravenous injection of synthetic human gastrin-17 in doses from 15.6 ng to 1 μg/kg increased the concentration of glucagon and insulin in peripheral venous blood to a maximum within 5 min. In spite of the enhanced concentrations of insulin induced by gastrin, corresponding concentrations of glucose were either unchanged or increased. Infusion of a mixture of fifteen aminoacids increased the concentrations of glucose, glucagon and insulin. While the increases in glucose and insulin concentrations were similar to those obtained after a protein-rich meal, the glucagon response was much larger after the infusion. Injection of gastrin-17 after 30 min of infusion of aminoacids did not potentiate either the glucagon or the insulin response.
The results indicate that gastrin, besides stimulating insulin secretion, can also stimulate glucagon secretion in a dose-dependent manner. The concentrations of gastrin necessary to stimulate glucagon secretion significantly correspond to the concentrations found in diseases with endogenous hypergastrinaemia (achlorhydria and Zollinger-Ellison syndrome). While gastrin potentiates the glucose-induced insulin secretion, it does not potentiate neither the aminoacid-induced insulin nor glucagon secretion.     

Progress in laparoscopic anatomy research: A review of the Chinese literature

The development of laparoscopic surgery has generated the new field of study, laparoscopic anatomy. This article reviews the reported literature on laparoscopic anatomy and explores how it has evolved along with advances in abdominal surgery. In addition, the principal concerns in current laparoscopic anatomy research are discussed, including: (1) types of special adjacent anatomical structures; and (2) special surgical planes and anatomical landmarks. Understanding of systematic laparoscopic anatomy can pr...     

Was lernen wir aus dem Fall Scott Reuben?

In February 2009 a major case of scientific misconduct was discovered. The American pain researcher Dr. S. Reuben had published 21 papers over a period of 15 years that were found to be fraudulent. Suddenly many advances in postoperative pain therapy which had been assumed to be correct seemed questionable. In this review article the lessons which can be learnt from this case are described. This review also reveals that it is almost impossible for reviewers or readers of scientific journals to detect scientific fraud. However, several warning signs can be identified that might be useful when reading clinical papers. In retrospect many of these signs were detectable in Reuben’s studies. Based on the fraudulent papers of Reuben it will be shown how and to what extent falsified results can affect other types of literature, such as practice guidelines, meta-analyses, review articles and oral presentations.     

Leukocytes as mediators of pain and analgesia

In inflammation, resident cells and infiltrating leukocytes produce proalgesic mediators. Although these mediators induce pain, the role of specific cell populations is still controversial. In addition, resident cells and leukocytes also generate analgesic mediators that counteract inflammatory pain, including anti-inflammatory cytokines, endocannabinoids, and opioid peptides. Chemokines and adhesion molecules orchestrate the migration of opioid peptide-containing leukocytes to inflamed tissue. Leukocytes secrete opioid peptides under stressful conditions or in response to releasing agents (eg, corticotropin-releasing factor and chemokines). Secretion requires intracellular calcium mobilization and activation of phosphinositol-3 kinase and p38 mitogen activated kinase. Following release, opioid peptides bind to receptors on peripheral sensory neurons and produce analgesia in animal models and humans. This review presents recent findings on the role of leukocytes in the generation and inhibition of inflammatory pain.     

Targeting of adenovirus vectors carrying a tumor cell-specific peptide: in vitro and in vivo studies

Previously, we have identified a tumor cell-specific peptide, HEW, by panning of phage display libraries on the human colorectal cancer cell line WiDr. In this report we demonstrate that this peptide can modify the infection properties of adenovirus vectors. Increased infectivity of replication-deficient adenovirus 5 vectors in WiDr cells was observed upon genetic insertion of the HEW peptide in the HI loop of the fiber knob. Moreover, whereas the coxsackie and adenovirus receptor (CAR)-ablating fiber mutation S408E abolished apparent infection in CAR-positive WiDr cells, the insertion of HEW completely restored infectivity toward these cells in vitro. To assess whether the de- and re-targeted infection profile was maintained in vivo, the fiber-modified adenovirus vectors were injected intratumorally or intravenously in WiDr tumor-bearing Swiss nu/nu mice. No significant differences in efficiency of infection could be observed suggesting alternative viral uptake mechanisms in vivo. Next, we have included the fiber shaft mutation S(*) in our studies, which was described to confer a de-targeted phenotype in vivo. Reduced gene transfer due to the S(*) mutation both in vitro and in vivo could be confirmed. Insertion of HEW in the HI knob loop of shaft-mutated fiber, however, did not rescue infectivity in target cells neither in vitro nor in vivo. We demonstrate the efficient ligand-mediated re-targeting of adenoviral vector infection to the human cancer cell line WiDr. The lack of apparent re-targeting in the in vivo situation is described.