Angiogenic potential of CD44+ CD90+ multipotent CNS stem cells in vitro

Although it is now clear that several subpopulations of neural stem cells (NSCs) exist during early development and adulthood, the angiogenic potential of NSCs remains a subject of debate. Here, we report that CD44(+) CD90(+) cells isolated from primary neurospheres can form vascular-tube structures in vitro. NSCs isolated from the mouse embryonic cortex formed neurospheres when cultured in serum-free medium containing 20ng/ml basic fibroblast growth factor (bFGF). CD44(+) CD90(+) cells were enriched from the neurospheres using an EPICS ALTRA flow cytometer, and antibodies against CD44 and CD90. The purified CD44(+) CD90(+) cells generated neurospheres, and differentiated into neurons and astrocytes. When the cells were inoculated into collagen gels and cultured with 20% fetal bovine serum plus bFGF for 7 days, vascular tube-like structures were formed. These results indicate that CD44(+) CD90(+) cells have the ability to generate neurospheres and to form vascular tubes.     

CD90 and CD110 correlate with cancer stem cell potentials in human T-acute lymphoblastic leukemia cells

Although cancer stem cells (CSCs) have been recently identified in myeloid leukemia, published data on lymphoid malignancy have been sparse. T-acute lymphoblastic leukemia (T-ALL) is characterized by the abnormal proliferation of T-cell precursors and is generally aggressive. As CD34 is the only positive-selection marker for CSCs in T-ALL, we performed extensive analysis of CD markers in T-ALL cell lines. We found that some of the tested lines consisted of heterogeneous populations of cells with various levels of surface marker expression. In particular, a small subpopulation of CD90 (Thy-1) and CD110 (c-Mpl) were shown to correlate with stem cell properties both in vitro and in transplantation experiments. As these markers are expressed on hematopoietic stem cells, our results suggest that stem cell-like population are enriched in CD90+/CD110+ fraction and they are useful positive-selection markers for the isolation of CSCs in some cases of T-ALL.     

Levels and values of circulating endothelial progenitor cells,soluble angiogenic factors,and mononuclear cell apoptosis in liver cirrhosis patients

Background

The roles of circulating endothelial progenitor cell (EPC) and mononuclear cell apoptosis (MCA) in liver cirrhosis (LC) patients are unknown. Moreover, vascular endothelial growth factor (VEGF) and stromal cell-derived factor (SDF)-1α are powerful endogenous substances enhancing EPC migration into circulation. We assessed the level and function of EPCs [CD31/CD34 (E₁), KDR/CD34 (E₂), CXCR4/CD34 (E₃)], levels of MCA, VEGF and SDF-1α in circulation of LC patients.

Methods

Blood sample was prospectively collected once for assessing EPC level and function, MCA, and plasma levels of VEGF and SDF-1α using flow cytometry and enzyme-linked immunosorbent assay (ELISA), respectively, in 78 LC patients and 25 age- and gender-matched healthy controls.

Results

Number of EPCs (E₁, E₂, E₃) was lower (all p < 0.0001), whereas SDF-1α level and MCA were higher (p < 0.001) in study patients compared with healthy controls. Number of EPCs (E₂, E₃) was higher but MCA was lower (all p < 0.05) in Child''s class A compared with Child''s class B and C patients, although no difference in VEGF and SDF-1α levels were noted among these patients. Chronic hepatitis B and esophageal varices bleeding were independently, whereas chronic hepatitis C, elevated aspartate aminotransferase (AST), and decompensated LC were inversely and independently correlated with circulating EPC level (all p < 0.03). Additionally, angiogenesis and transwell migratory ability of EPCs were reduced in LC patients than in controls (all p < 0.001).

Conclusion

The results of this study demonstrated that level, angiogenic capacity, and function of circulating EPCs were significantly reduced, whereas plasma levels of SDF-1α and circulating MCA were substantially enhanced in cirrhotic patients.     

The role of the CD39–CD73–adenosine pathway in liver disease

Extracellular adenosine triphosphate (ATP) is a danger signal released by dying and damaged cells, and it functions as an immunostimulatory signal that promotes inflammation. The ectonucleotidases CD39/ectonucleoside triphosphate diphosphohydrolase‐1 and CD73/ecto‐5′‐nucleotidase are cell‐surface enzymes that breakdown extracellular ATP into adenosine. This drives a shift from an ATP‐driven proinflammatory environment to an anti‐inflammatory milieu induced by adenosine. The CD39–CD73–adenosine pathway changes dynamically with the pathophysiological context in which it is embedded. Accumulating evidence suggests that CD39 and CD73 play important roles in liver disease as critical components of the extracellular adenosinergic pathway. Recent studies have shown that the modification of the CD39–CD73–adenosine pathway alters the liver's response to injury. Moreover, adenosine exerts different effects on the pathophysiology of the liver through different receptors. In this review, we aim to describe the role of the CD39–CD73–adenosine pathway and adenosine receptors in liver disease, highlighting potential therapeutic targets in this pathway, which will facilitate the development of therapeutic strategies for the treatment of liver disease.     

Hepatic stem cells: A viable approach for the treatment of liver cirrhosis

Liver cirrhosis is characterized by distortion of liver architecture, necrosis of hepatocytes and regenerative nodules formation leading to cirrhosis. Various types of cell sources have been used for the management and treatment of decompensated liver cirrhosis. Knowledge of stem cells has offered a new dimension for regenerative therapy and has been considered as one of the potential adjuvant treatment modality in patients with end stage liver diseases (ESLD). Human fetal hepatic progenitor cells are less immunogenic than adult ones. They are highly propagative and challenging to cryopreservation. In our earlier studies we have demonstrated that fetuses at 10-18 wk of gestation age contain a large number of actively dividing hepatic stem and progenitor cells which possess bi-potent nature having potential to differentiate into bile duct cells and mature hepatocytes. Hepatic stem cell therapy for the treatment of ESLD is in their early stage of the translation. The emerging technology of decellularization and recellularization might offer a significant platform for developing bioengineered personalized livers to come over the scarcity of desired number of donor organs for the treatment of ESLD. Despite these significant advancements long-term tracking of stem cells in human is the most important subject nowadays in order to answer several unsettles issues regarding the route of delivery, the choice of stem cell type(s), the cell number and the time-point of cell delivery for the treatment in a chronic setting. Answering to these questions will further contribute to the development of safer, noninvasive, and repeatable imaging modalities that could discover better cell therapeutic approaches from bench to bed-side. Combinatorial approach of decellularization and nanotechnology could pave a way towards the better understanding in determination of cell fate post-transplantation.     

Isolation and characterization of parenchymal cells from experimentally induced macronodular rat liver cirrhosis

Hepatocytes were isolated from thioacetamide (TAA)-induced macronodular cirrhotic rat livers by a collagenase perfusion method. In the content of cellular metabolites, fatty acid uptake and lipid secretion there were no substantial differences compared with cells isolated from micronodular cirrhosis described previously. In contrast to isolated hepatocytes from normal livers those from macronodular cirrhosis had a lowered cellular content of triglycerides, phospholipids and cholesterol but not of cholesterol esters and free fatty acids. In macronodular cirrhosis hepatocytes of hypertrophic type, rich in cell organelles, can be distinguished ultrastructurally from those with signs of atrophy and degeneration. Immediately after isolation many hepatocytes isolated from macronodular cirrhosis showed plasma membrane blebbing. Whereas the blebbing was without recognizable effects on the fine structure of the isolated hepatocytes of the hypertrophic type, in the more atrophic ones some mitochondria were swollen. In addition, morphological analysis of the crude and purified suspensions revealed a partial selection of the hypertrophic cells during the isolation procedure, presumably due to a more labile state of those cells which showed signs of atrophy and degeneration. When stabilized in the suspension medium, however, the hepatocytes maintained complex metabolic functions for at least 2 h. Thus, the method described allows the isolation of parenchymal cells from TAA-induced macronodular cirrhotic livers for studying ultrastructural and biochemical alterations in hyperregenerative experimental liver cirrhosis.     

Golgi protein 73 and its diagnostic value in liver diseases

Golgi protein 73 (GP73, also referred to as Golph 2) with 400 amino acids is a 73 kDa transmembrane glycoprotein typically found in the cis‐Golg complex. It is primarily expressed in epithelial cells, which has been found upregulated in hepatocytes in patients suffering from both viral and non‐viral liver diseases. GP73 has drawn increasing attention for its potential application in the diagnosis of liver diseases such as hepatitis, liver cirrhosis and liver cancer. Herein, we reviewed the discovery history of GP73 and summarized studies by many groups around the world, aiming at understanding its structure, expression, function, detection methods and the relationship between GP73 and liver diseases in various settings.     

Therapeutic benefits of CD90‐negative cardiac stromal cells in rats with a 30‐day chronic infarct

Cardiac stromal cells (CSCs) can be derived from explant cultures, and a subgroup of these cells is viewed as cardiac mesenchymal stem cells due to their expression of CD90. Here, we sought to determine the therapeutic potential of CD90‐positive and CD90‐negative CSCs in a rat model of chronic myocardial infarction. We obtain CD90‐positive and CD90‐negative fractions of CSCs from rat myocardial tissue explant cultures by magnetically activated cell sorting. In vitro, CD90‐negative CSCs outperform CD90‐positive CSCs in tube formation and cardiomyocyte functional assays. In rats with a 30‐day infarct, injection of CD90‐negative CSCs augments cardiac function in the infarct in a way superior to that from CD90‐positive CSCs and unsorted CSCs. Histological analysis revealed that CD90‐negative CSCs increase vascularization in the infarct. Our results suggest that CD90‐negative CSCs could be a development candidate as a new cell therapy product for chronic myocardial infarction.     

CD73在临床疾病中的研究进展

CD73又称5'-核苷酸酶,普遍存在人体多种细胞,是通过糖基-磷酰肌醇介导连接在胞膜上的糖蛋白。CD73有水解酶活性,可降解5'-磷酸腺苷成腺苷,进而通过腺苷与各种腺苷受体亚型作用发挥血管生成、旁分泌和免疫抑制等作用;此外CD73可发挥非酶作用,介导细胞间结合和信号传递。CD73与肿瘤、心肌损伤、脑损伤、糖尿病、弓形虫入侵、系统性红斑狼疮和器官移植等多种临床疾病相关,影响疾病的发生发展,CD73及其水解产生的腺苷与腺苷受体作用,影响机体的生理和病理过程。本文综述近年来CD73的基础和临床研究,以明晰CD73基础研究和临床应用的联系,加快CD73的临床应用。     

肝硬化合并结核病患者的临床特征

目的 分析肝硬化合并结核病患者的临床特征,为该类患者的治疗提供参考。 方法 回顾性分析2014年9月至2017年7月于浙江大学医学院附属第一医院住院的肝硬化合并结核病患者的相关资料,总结患者的临床特征。 结果 共收治68例肝硬化合并结核病患者,其中男性50例(73.5%),女性18例(26.5%),平均年龄(59.9±13.7)岁。在所有患者中,肝硬化最常见的原因是HBV感染(46例,67.7%)和酒精(9例,13.2%)。肺结核(43例)、结核性腹膜炎(13例)、结核性胸膜炎(12例)是最常见的结核类型,此外骨关节结核、结核性脑膜炎、结核性心包炎、泌尿系结核、肠结核、淋巴结结核均可见。该类患者常见临床表现为乏力、腹胀,出现发热的患者不足半数,有盗汗表现的更为罕见。 结论 肝硬化患者并发结核后可使病情复杂化,增加治疗难度。此外部分患者临床表现不典型,需提高警惕,避免漏诊。     

CD90-positive cells, an additional cell population, produce laminin alpha2 upon transplantation to dy(3k)/dy(3k) mice

Laminin alpha2 is a component of skeletal and cardiac muscle basal lamina. A defect of the laminin alpha2 chain leads to severe congenital muscular dystrophy (MDC1A) in humans and dy/dy mice. Myogenic cells including myoblasts, myotubes, and myofibers in skeletal muscle are a possible source of the laminin alpha2 chain, and myogenic cells are thus proposed as a cell source for congenital muscular dystrophy therapy. However, we observed production of laminin alpha2 in non-myogenic cells of normal mice, and we could enrich these laminin alpha2-producing cells in CD90(+) cell fractions. Intriguingly, the number of CD90(+) cells increased dramatically during skeletal muscle regeneration in mice. This fraction did not include myogenic cells but exhibited a fibroblast-like phenotype. Moreover, these cells were resident in skeletal muscle, not derived from bone marrow. Finally, the production of laminin alpha2 in CD90(+) cells was not dependent on fusion with myogenic cells. Thus, CD90(+) cells are a newly identified additional cell fraction that increased during skeletal muscle regeneration in vivo and could be another cell source for therapy for lama2-deficient muscular dystrophy.     

CD90过表达影响胃癌细胞AGS的干性特征

2019年中国癌症报告显示,胃癌发病率仅次于肺癌,位列第二,其死亡率排在所有肿瘤的第三位,严重危害人们健康.筛查和鉴定胃癌的早期检测标志物、寻找胃癌治疗的分子靶点,对于降低胃癌致死率至关重要.CD90 (THY1)是一种细胞表面糖蛋白,在肿瘤细胞增殖、转移以及血管生成中发挥重要作用.CD90异常表达与干细胞特性有关,促...     

肝硬化患者肠道菌群失衡的研究现状

肝硬化是我国的一种常见病,近年来越来越多的研究表明肝硬化及其并发症（如门静脉高压、自发性腹膜炎、肝性脑病及肝癌等）都与肠道菌群失衡有着密切的联系。肝脏和肠道通过“肠—肝轴”紧密联系在一起,肝硬化时因小肠细菌过度生长、肠黏膜屏障功能受损、机体免疫功能下降等因素,导致细菌移位、肠道微生态失衡。而肠道微生态失衡又会使肝功能障碍进一步发展,引起肝性脑病等并发症。本文就目前国内外对肝硬化及其并发症与肠道细菌及真菌菌群失衡的研究进行综述。     

HCV-specific interleukin-21+CD4+ T cells responses associated with viral control through the modulation of HCV-specific CD8+ T cells function in chronic hepatitis C patients

Interleukin-21 (IL-21)+CD4+ T cells are involved in the immune response against hepatitis B virus (HBV) by secreting IL-21. However, the role of IL-21+CD4+ T cells in the immune response against chronic hepatitis C (CHC) virus infection is poorly understood. This study aimed to investigate the role of IL-21+CD4+ T cells in CHC patients and the potential mechanisms. The study subjects included nineteen CHC patients who were grouped by viral load (low, < 10⁶ RNA copies/ml, n = 8; high, > 10⁶ RNA copies/ml, n = 11). The peripheral frequency of HCV-specific IL-21+CD4+ T cells was higher in the low viral load group and was negatively correlated with the serum HCV RNA viral load in all CHC patients. Meanwhile, IL-21+ cells accumulated in the liver in the low viral load group. In vitro, IL-21 treatment increased the expression of proliferation markers and cytolytic molecules on HCV-specific CD8+ T cells. In summary, these findings suggest that HCV-specific IL-21+CD4+ T cells might contribute to HCV control by rescuing HCV-specific CD8+ T cells in CHC patients.     

肝纤维化基因治疗的进展

肝硬化是慢性肝病晚期的组织学改变 ,以纤维组织大量增生和肝小叶结构无序化为特征 ,因此又称肝纤维化。近年来随着分子生物学的发展 ,肝纤维化的分子机制逐渐得以阐明 ,从而使肝纤维化的基因治疗成为可能。肝纤维化的基因治疗主要起到阻止纤维化发展、刺激肝细胞分裂和肝组织结构重建三方面的作用。目前 ,常用的方法一般是通过缺陷病毒 (如腺病毒 )转入特定的细胞因子和酶 (如肝细胞生长因子、转化生长因子β1受体、基质金属蛋白酶等 )的基因 ,通过靶细胞表达这些因子作用于受损的肝脏 ,达到延缓和治愈肝纤维化的目的     

复合益生菌活菌制剂对肝硬化患者肠道菌群的影响

目的:观察肝硬化患者口服三联活菌后肠道菌群、血氧及血浆内毒素的变化.方法:选择肠道菌群中具有代表性的细菌进行培养和计数.42例肝硬化患者给予三联活菌治疗21 d.测定治疗前后肠道菌群菌落计数、血氨(干片法)、血浆内毒素(改良鲎试验法).结果:与正常对照组相比,肝硬化组患者存在不同程度的肠道菌群失调,主要表现为双歧杆菌减少(10.12±0.71比9.27±1.25,P＜0.05).治疗后,双歧杆菌由9.27±1.25增至10.43±1.25,差异有显著性(P＜0.01).且血氨水平降低,并可降低肝硬化患者血浆内毒素水平[(109.21±12.23)pg/ml比(71.46±9.12)pg/ml,P＜0.05].结论:肝硬化患者存在肠道菌群失调.益生菌制剂可有效改善肝硬化患者肠道菌群失调,并降低血氨及血浆内毒素.     

Influence of NSAIDs and methotrexate on CD73 expression and glioma cell growth

Glioblastoma (GBM) is the most malignant and deadly brain tumor. GBM cells overexpress the CD73 enzyme, which controls the level of extracellular adenosine, an immunosuppressive molecule. Studies have shown that some nonsteroidal anti-inflammatory drugs (NSAIDs) and methotrexate (MTX) have antiproliferative and modulatory effects on CD73 in vitro and in vivo. However, it remains unclear whether the antiproliferative effects of MTX and NSAIDS in GBM cells are mediated by increases in CD73 expression and adenosine formation. The aim of this study was to evaluate the effect of the NSAIDs, naproxen, piroxicam, meloxicam, ibuprofen, sodium diclofenac, acetylsalicylic acid, nimesulide, and ketoprofen on CD73 expression in GBM and mononuclear cells. In addition, we sought to understand whether the effects of MTX may be mediated by CD73 expression and activity. Cell viability and CD73 expression were evaluated in C6 and mononuclear cells after exposure to NSAIDs. For analysis of the mechanism of action of MTX, GBM cells were treated with APCP (CD73 inhibitor), dipyridamole (inhibitor of adenosine uptake), ABT-702 (adenosine kinase enzyme inhibitor), or caffeine (P1 adenosine receptor antagonist), before treatment with MTX and AMP, in the presence or not of mononuclear cells. In summary, only MTX increased the expression of CD73 in GBM cells decreasing cells viability by mechanisms independent of the adenosinergic system. Further studies are needed to understand the role of MTX in the GBM microenvironment.

     

Isolation and characterization of parenchymal cells from normal and cirrhotic rat liver

A technique is described for isolation of adult rat hepatocytes from micronodular cirrhotic livers based on a collagenase digestion procedure. Hepatocytes from normal livers and those chronically injured by thioacetamide did not differ with respect to the viability measured by the trypan blue exclusion test or to the cellular concentrations of protein and glycogen, but the triglyceride content of cells from cirrhotic livers was significantly reduced. Hepatocytes isolated from cirrhotic livers are ultrastructurally in a good state of preservation but they appear to be poorer than controls in RER membranes, although the well-preserved mitochondria are somewhat richer in cristae. No differences were detected between the cell preparations in rates of gluconeogenesis and total de novo fatty acid synthesis, but the secretion of newly synthesized fatty acids was significantly reduced in cells from cirrhotic livers. Thus adult rat hepatocytes can be isolated from thioacetamide-induced micronodular cirrhotic livers with high yield and morphological integrity. Differentiated functions are maintained in suspension for at least 4 h.