肝细胞的三维受控组装

为解决肝病救治中肝供体缺乏的问题,采用基于快速成形技术的细胞组装机,将肝细胞和海藻酸钠与明胶复合材料的共混物作为成形对象,通过细胞材料直接三维受控组装技术,实现了堆积成形具有一定三维结构和预定义孔隙的肝组织前体。常规体外培养条件下,肝细胞存活并保持生物学活性达12d之久。这一技术有望成为肝脏及其他人体组织器官人工构造的新途径。     

酒精性肝病患者瘦素水平与肝功能的相关性

目的 探讨瘦素在酒精性肝病(ALD)及不同阶段中的变化规律及其与肝功能受损的关系.方法 采用放射免疫法分别测定22例酒精性脂肪肝(AFD)、11例酒精性肝炎(AH)、13例酒精性肝硬化(AC)患者的血清瘦素水平,同时生化检测46例患者的肝功能,选择30例健康体检者作对照.结果 酒精性肝病患者血清瘦素水平(10.63±4.06 μg/L)明显高于正常对照组(4.98±2.89 μg/L),P<0.05,酒精性肝病患者血清瘦素水平与肝功能受损呈明显相关性.结论 测定血清瘦素水平可用于评估酒精性肝病的病情,对判断预后有参考价值.     

Synchrotron refractive-index microradiography of human liver cancer tissue

Three human liver tissue samples （-5 mm × 40 mm × 20 mm） were excised from a cancer patient＇s liver during surgery. The microradiology analysis was performed with a non-standard approach on a synchrotron. High-resolution refractive-index edge-enhanced microradiographs that cover a larger volume of the liver tissue sample were obtained. The cancer tissue and normal tissue could be clearly identified and distinguished based on their different textures. Furthermore, new blood vessel hyperplasia was found near the cancer area. Blood vessels with a diameter smaller than 20 μm could be identified. These findings were fully consistent with the histopathological examination or the same area. Microradiographs of the newly formed blood vessels at different angles were also obtained. This result shows that it is possible to further develop this approach into a technique of microradiographic imaging for clinic diagnosis of liver cancer at the early stage.     

原位辅助性部分肝移植

 自1989年原位辅助性部分肝移植成功应用于临床治疗急性肝功能衰竭以来,适应证逐渐扩大至肝功能衰竭、遗传代谢疾病、终末期肝硬化。随技术不断进步趋于成熟,并发症发生率显著降低,远期存活率明显上升,对部分疾病治疗效果达到甚至优于原位肝移植术,并衍生出多米诺原位辅助性部分肝移植等新术式,显著拓展了供体来源及受益人群,成为肝脏疾病重要的治疗手段。本文介绍原位辅助性部分肝移植术的发展历程及研究进展。     

Therapeutic applications of bone marrow-derived stem cells in liver transplantation for end-stage liver diseases

Today, liver transplantation (LT) is the only established treatment for end-stage liver diseases. The de- velopment of LT, including OLT, cadaveric LT, split LT, living donor LT (LDLT), brings hopes to patients with these diseases. However, increasing donor shortage, rejection and life-long immunosuppression with its side effects are the major limitations of this therapy strategy. Bone marrow-derived stem cells (BMDSCs) are capable of differentiating into hepatocyte-like cells and contribute to liver injury repair. The microenvironment of liver injury caused by rejection, ischemia/reperfusion, loss of liver mass, recurrence of HCV and "small-for-size syndrome" after LT can attract a variety of bone marrow-derived stem cell population to the peripheral circulation and then migration to the injury liver to promote the hepatic function restoration. Additionally, BMDSCs can also take part in the functional regeneration of living donor liver after LDLT. This participation in liver regeneration may be associated to the interac- tion between SDF-1and its receptor CXCR4, involving HGF, IL-8, MMP9, and VEGF/VEGFR-2. BMDSC with its bio-characteristics could maintain the allograft tolerance from different angles and in different ways. In conclusion, BMDSCs transplantation, as a new assistant therapeutic method for LT, will ex- pand the space of LT, and provide more survival opportunities for the patients suffering liver diseases in the future.     

基于模糊神经网络的产品可装配性评判

针对产品装配信息量大、影响因素复杂、许多参数依赖人的判断和推理等特点,以及多数现行评判方法存在的不足,提出了基于模糊神经网络的产品可装配性评判方法·该方法依据产品装配特征的类型,先构建产品可装配性信息数据库,再通过将常规的三层BP网络模糊化处理,得到输入和权值均已模糊化的产品可装配性模糊神经网络模型·按照预置的网络训练规则进行训练,获得最佳连接权值·经实例检验,证明这种模糊神经网络模型用于产品可装配性评判是可行的,且具有很好的稳定性·     

肝硬化肝切除术后肝再生及评估指标研究

原发性肝癌是我国的常见病、多发病,且90%的患者合并有肝硬化,肝切除术是主要的治疗方法,而肝脏在受到损害或切除之后,可以显示出强大的再生能力,这一现象早已被人们所发现并研究.就肝硬化肝切除术后肝再生、肝再生受损机制以及肝再生评估予以综述.     

小鼠胚肝基质细胞在体外培养中的动态演化

胚肝基质细胞是研究胚肝造血调控的重要工具，本实验对小鼠胚肝基质细胞在体外培养过程中的生长、细胞类型组成及其组成比例的演变过程进行了观察．实验显示小鼠原代培养初期基质细胞是由平滑肌样上皮细胞、巨噬细胞以及成纤维样细胞、树突状细胞、内皮细胞构成的，经过多次传代后，巨噬细胞等类型的细胞逐渐丢失，平滑肌样上皮细胞和成纤维样细胞成为主体，表明体外培养的小鼠胚肝基质细胞在4代以内都能较真实的反映体内造血微环境的构成，适用于胚肝造血研究．此为进一步研究胚肝基质细胞在胚胎期造血过程中的作用奠定了工作基础．     

肝癌临床标本原位移植模型的建立及其影像学评估

目的 建立基于临床肝癌手术标本的原位移植(patient-derived orthotopic xenograft PDOX)模型,研究近红外荧光(Near-infrared fluorescence,NIRF)活体成像和PET/CT在模型评估中的应用。方法 将临床新鲜肝癌手术标本接种于重度免疫缺陷NPG小鼠皮下建立肝癌PDX模型,通过组织形态观察、STR分型检测和免疫组化分析对PDX模型进行评估。进一步将PDX模型肿瘤组织进行裸鼠肝原位移植建立PDOX模型,注射近红外荧光染料IR-783,通过小动物活体成像检测肿瘤的发生;尾静脉注射18F-FDG,通过小动物PET/CT观察确认肝原位肿瘤的生长。结果 STR分型结果表明PDX肿瘤的人源性特征,组织形态观察和免疫组化检测表明PDX肿瘤保持了原发肿瘤病理学特征;近红外荧光活体成像检测到肝脏肿瘤的发生;PET/CT可清晰观察到小鼠肝脏部位18F-FDG分子探针富集。结论 成功建立了肝癌PDOX模型,通过小动物活体成像和PET/CT影像技术可对该模型进行评估,为肝癌的治疗和发病机制研究提供了良好的动物模型。     

筛选hALR的相互作用蛋白基因

为筛选与人肝再生增强因子(hALR)相互作用蛋白的基因，探讨肝再生增强因子在肝再生过程中的分子生物学机制，将人肝再生增强因子基因的开放读码框片断，重组入载体pGBKT7构建成“诱饵”质粒pGBKT7—hALR，然后用酵母双杂交系统从预转化酵母菌Y187的成人肝细胞cDNA文库中筛选与人肝再生增强因子相互作用蛋白的基因。筛出的阳性克隆基因序列被测出后，经GenBank查询，结果发现它们分别是血清白蛋白、金属硫蛋白、硒蛋白类似物、Na^ ／K^ ATPase和一个未知功能的蛋白的部分序列。这初步克隆了与hALR相互作用的蛋白基因，为进一步探讨ALR在肝再生过程中的作用机制奠定了基础。     

筛选hALR的相互作用蛋白基因(英)

为筛选与人肝再生增强因子(hALR)相互作用蛋白的基因,探讨肝再生增强因子在肝再生过程中的分子生物学机制,将人肝再生增强因子基因的开放读码框片断,重组入载体pGBKT7构建成“诱饵”质粒pGBKT7-hALR,然后用酵母双杂交系统从预转化酵母菌Y187的成人肝细胞cDNA文库中筛选与人肝再生增强因子相互作用蛋白的基因.筛出的阳性克隆基因序列被测出后,经GenBank查询,结果发现它们分别是血清白蛋白、金属硫蛋白、硒蛋白类似物、Na+/K+ ATPase和一个未知功能的蛋白的部分序列.这初步克隆了与hALR相互作用的蛋白基因,为进一步探讨ALR在肝再生过程中的作用机制奠定了基础.     

面向复杂产品交互虚拟装配操作的并行碰撞检测算法

运动对象间碰撞检测是交互式虚拟装配的一个基本问题,提出一种虚拟环境中运动对象间的并行化碰撞检测方法.该方法使用一种并行的、基于区域分割和快速相交校验排序的分解算法来包围盒层次模型,检测计算时依据用户的操作动态决定碰撞检测对以减少检测计算量,同时基于微机和局域网的并行方法来计算模型间碰撞,两个模型间碰撞检测时使用包围盒层次树动态更新方法.随后,以某型汽车底盘虚拟装配时的实时碰撞检测来验证算法性能.结果表明,该方法可以快速建立包围盒层次树模型,并可在交互操作中完成给定精度的实时碰撞检测.     

蜂窝CDMA系统多区基站天线倾斜方案

在综合考虑整个蜂窝系统的情况下,结合环境阴影衰落深度等因素,分析了地面基站天线倾斜对CDMA系统整体容量的影响后,提出了一种CDMA系统多区基站的天线倾斜方案.该方案将基站天线倾斜技术应用于整个CDMA蜂窝系统.仿真结果表明,利用本方法可将CDMA系统容量得到明显的提高.     

VG染色对慢性肝病肝纤维化的研究

目的：应用组织化学染色对６１例肝活检标本进行研究,观察胶原纤维在慢性肝病组织中的分布,研究由慢性肝炎到肝硬化肝纤维化的形成过程。方法：应用ＶＧ染色显示慢性肝病组织中的胶原纤维,光镜观察。结果：胶原纤维染成粉红色,形成间隔分割肝实质。结论：ＶＧ染色对慢性肝病有诊断学意义。肝实质内的纤维组织可能来源于成纤维细胞和贮脂细胞。     

Construct hepatic analog by cell-matrix controlled assembly technology

Living donor liver transplantation has been widely performed since 1989[1]. Unfortunately, the demand for donor livers far outstrips the supply. The severe short- age of donor livers continues to grow, thereby stimu- lating people to develop alternate eff…     

Subcellular localization of several heavy metals of Hg, Cd and Pb in human liver

Liver, as an important metabolic and detoxicological organ of human body, can be used as a good bioindicator for evaluating body burden of environmental pollutants. Its elemental contents and their chemical forms are closely related to the status of human health and disease. In this paper, the liver samples collected from normal subjects were separated to different subcellular fractions of nuclei, mitochondria, lysosome, microsome and cytosol by differential centrifugation. Then their concentrations of heavy metals of As, Pb, Cd, and Hg were determined by atomic absorption and atomic fluorescent spectroscopy. Our results show no significant difference with literature ones when comparing their gross concentrations. In the case of their subcellular distribution, the Hg concentrations are higher in mitochondrial, microsomal and cytosolic fractions; the Cd concentrations are higher in cytosolic and mitochondrial fractions, while As highest in nuclear fraction. The highest concentration of Pb is found in microsomal fraction with similarity to Fe. Mercury in liver is mainly in the form of inorganic, and methylmercury ranged from 9% to 50% with the average value of 20.9% 13.3%. These results indicate that the cellular distribution and the accumulated target organelles are quite different among these heavy metals, which suggest their various pathways and toxic mechanism in vivo.     

Two-dimensional gel electrophoresis analysis of the proteomes expressed in the human hepatoma cell line BEL-7404 and normal liver cell line L-02

Proteome analysis technology has been used extensively in conducting discovery research of biology and has become one of the most essential technologies in functional genomics. The proteomes of the human hepatoma cell line BEL-7404 and the normal human liver cell line L-02 have been separated by high resolution two-dimensional gel electrophoresis (2-DE) with immobilized pH gradient isoelectric focusing (IPG-IEF) in the first dimension and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) in the second dimension (IPG-DALT). The resulting images have been analyzed using 2-D analysis software. Quantitative analysis reveals that 7 protein spots are detected only in hepatoma BEL-7404 cells, 14 only in L-02 cells, and 78 protein spots show significant fluctuation in quantity in both cell lines (P< 0.01). These protein spots have been displayed on a proteome differential expression map. Analysis for the reproducibility of 2-DE indicates that the positional variability in the IEF dimension is 0.73 mm, while the variability in the SDS-PAGE dimension is 0.44 mm, and the quantitative variability is 17.6%–19.2%. These results suggest that the reproducibility of 2-DE has been suitable for the study of differential expression of proteomes. Proteome differential expression maps can be useful tools for disease diagnosis, drug-target validation analysis and biological process elucidation.     

Differentiation of human embryonic stem cells along a hepatocyte lineage and its application in liver regeneration

Hepatocyte transplantation and bioarUficial liver （BAL） as alternatives to liver transplantation offer the possibility of effective treatment for many inherited and acquired hepatic disorders. Unfortunately, the limited availability of donated livers and the variability of their derived hepatocytes make it difficult to obtain enough viable human hepatocytes for the hepatocyte-based therapies. Embryonic stem cells （ESCs）, which could be isolated directly from the blastocyst inner cell mass, have permanent self-renewal capability and developmental pluripotency and therefore might be an ideal cell source in the treatment of hepatic discords. However, differentiation of hESCs into hepatocytes with significant numbers remains a challenge. This review updates our current understanding of differentiation of ESCs into hepatic lineage cells, their future therapeutic uses and problems in liver regeneration.     

输注体外诱导的同种反应性淋巴细胞诱导大鼠肝脏移植免疫耐受的研究

目的 观察输注体外诱导的同种反应性淋巴细胞,是否可以延长大鼠移植肝脏的存活时间,从而探索一条诱导肝移植免疫耐受的新途径.方法 以供者Lewis大鼠淋巴细胞作为刺激物,体外刺激受者BN大鼠淋巴细胞增殖,应用CFSE（羧基荧光素二醋酸盐琥珀酰亚胺酯）标记法检测增殖情况;将增殖后的BN大鼠淋巴细胞经照射灭活后输注给同品系的BN大鼠,并行Lewis到BN的肝移植,观察肝移植后BN大鼠生存期.结果 同种反应性淋巴细胞体外能够活跃增殖;采用同种反应性淋巴细胞输注可以显著延长肝移植大鼠的存活时间（P＜0.05）.结论 体外诱导的同种反应性淋巴细胞输注,有可能成为诱导抗原特异性免疫耐受的一种新途径.     

New approach to paricalcitol synthesis

Paricalcitol,an analog of vitamin D,is used as a drug for the prevention and treatment of secondary hyperparathyroidism.In this paper,a new strategy for the synthesis of paricalcitol is described.This approach includes three main improvements:one-pot regioselective ozonization cleavage of the side-chain and methylene at C-19,free-radical reduction removal of the OH group formed at C-19,and side-chain assembly using a Wittig reaction.These are all new strategies for the synthesis of 19-nor-vitamin D2 compounds.