猪作为异种器官移植供体的研究进展

异种器官移植(Xenotransplantation,XT)是未来生物学科中的重要研究领域之一.猪作为为人类提供移植器官的供体源具有得天独厚的条件.文章就器官移植(OrganTransplantation,OT)的历史、原因、移植后的免疫排斥反应和猪内源性逆转录病毒(porcine endogenous retrovirous,PERV)感染问题及可能的解决途径进行了综述.     

临床异种移植百年进展(述评)

美国最近连续报道4例基因工程猪供给人的异种器官移植，这些进展打破了异种移植临床研究领域近30年的沉静。尤其是实验性的猪心脏移植取得初步成功，引发了广泛的关注。异种移植面向未来的临床应用迈出了关键性一步。这4个案例，无论是创新精神、技术进步、审批流程，还是伦理学的同步跟进，都值得我国同行学习和借鉴。30年来，基因工程（GE）动物器官已经能在非人类灵长类动物（NHPs）模型中长时间存活。这些实验中，预存抗体、补体激活、凝血状态等机制参与的超急性排异反应基本上得到有效控制。异种移植已经达到临床应用的起跑点。然而，未来仍然有四大问题需要解决：免疫排斥、生物安全、跨种适配和伦理心理。本文简要概括临床异种移植百年来的主要进展，其困难程度可见一斑，但希望和探索从未终止。     

HLA－G inhibits xenogenetic cytotoxicity mediated by human NK cells and T lymphocytes against PECs

In order to investigate whether the non-classical HLA-G class I molecule protects the prcine endothelial cells(PECs)from the lysis mediated by human immune cells in pig to human discordant xenotransplantation,we have cloned HLA-G cDNA from a human placents by RT-PCR.Mammalian expression vector,pEFG-neo,was constructed by insertion of HLA-G cDNA in pEF-neo.We obtained efficiently expressed PECs by stable transfection.Cytotoxicity assay showed that overexpression of HLA-G on PECs was sufficient to inhibit human NK-92 cell lysis.The level of lysis was equal to or less than that of the lysis of human umbilical vein endothelial cells mediated by human NK-92 cells.It also indicated that HLA-G inhibited the lysis of PECs mediated by xeno-antigen specific T lymphocytes.The reduction of lysis ranged between 59.1% and 88.9A%.These findings suggest that the transgenic approach to overexpress HLA-G is believed to be a new immunotherapy in overconing the immune rejections in xenotransplantion,including delayed xenograft rejection and cell-mediated rejection.     

HLA-G inhibits xenogenetic cytotoxicity mediated by human NK cells and T lymphocytes against PECs

The worldwide shortage in the supply of human do-nor organs is becoming more and more pronounced. Xenotransplantation may probably give the hope to over-come the problem ultimately. Because it has a great pros-pect of clinical application, xenotransplantation has drawn great attention[1]. The pig appears to be an ideal source for human transplantation. But the xenograft has to face the challenge of three severe rejections (the hyperacute rejec-tion, the delayed xenograft rejection and acute ce…     

我国当前重要人体食源性寄生虫病原

为考察我国当前重要人体食源性寄生虫病原的种类、新的流行感染方式和感染途径．本文论述了我国近几年出现的重要食源性人兽共患寄生虫病原种类，介绍病原的感染来源、感染途径和感染方式．结果经统计发现有29种食源性寄生虫病原．这些寄生虫病原的感染来源主要通过日常食物污染，经口随食物感染，并且多数为人兽共患寄生虫病原．     

Identification of cells initiating human melanomas

Tumour-initiating cells capable of self-renewal and differentiation, which are responsible for tumour growth, have been identified in human haematological malignancies and solid cancers. If such minority populations are associated with tumour progression in human patients, specific targeting of tumour-initiating cells could be a strategy to eradicate cancers currently resistant to systemic therapy. Here we identify a subpopulation enriched for human malignant-melanoma-initiating cells (MMIC) defined by expression of the chemoresistance mediator ABCB5 (refs 7, 8) and show that specific targeting of this tumorigenic minority population inhibits tumour growth. ABCB5+ tumour cells detected in human melanoma patients show a primitive molecular phenotype and correlate with clinical melanoma progression. In serial human-to-mouse xenotransplantation experiments, ABCB5+ melanoma cells possess greater tumorigenic capacity than ABCB5- bulk populations and re-establish clinical tumour heterogeneity. In vivo genetic lineage tracking demonstrates a specific capacity of ABCB5+ subpopulations for self-renewal and differentiation, because ABCB5+ cancer cells generate both ABCB5+ and ABCB5- progeny, whereas ABCB5- tumour populations give rise, at lower rates, exclusively to ABCB5- cells. In an initial proof-of-principle analysis, designed to test the hypothesis that MMIC are also required for growth of established tumours, systemic administration of a monoclonal antibody directed at ABCB5, shown to be capable of inducing antibody-dependent cell-mediated cytotoxicity in ABCB5+ MMIC, exerted tumour-inhibitory effects. Identification of tumour-initiating cells with enhanced abundance in more advanced disease but susceptibility to specific targeting through a defining chemoresistance determinant has important implications for cancer therapy.     

基于卷积神经网络的猪脸特征点检测方法

针对牲畜面部识别在养殖行业广泛需求的问题, 提出一种基于卷积神经网络的猪脸特征点检测方法, 解决了猪脸特征点难检测的问题. 首先, 采集猪面部数据并进行特征点标注, 使用新的采集方法以解决猪口部通常不可见的问题； 其次, 对猪脸数据和人脸数据进行结构计算, 匹配相似度较高的猪脸和人脸, 构建猪脸人脸匹配数据集； 再次, 利用匹配数据集训练TPS(thin plate spline)形变卷积神经网络, 得到形变后的猪脸数据集以适配人脸特征点检测模型； 最后, 使用形变猪脸数据集对人脸特征点检测神经网络模型进行微调, 得到猪脸特征点检测模型. 实验结果表明, 用该方法进行猪脸特征点检测, 错误率仅为5.60%.     

HIV susceptibility conferred to human fibroblasts by cytomegalovirus-induced Fc receptor

The main receptor for the human immunodeficiency viruses type 1 and 2 (HIV-1 and HIV-2) on T and B lymphocytes, monocytes and macrophages is the CD4 antigen 1-3. Infection of these cells is blocked by monoclonal antibodies to CD4(1,2) and by recombinant soluble CD4(4-9). Expression of transfected CD4 on the surface of HeLa and other human cells renders them susceptible to HIV infection 10. HIV-antibody complexes can also infect monocytes and macrophages by means of receptors for the Fc portion of immunoglobulins (FcR)11-13), or complement receptors 14,15. The expression of IgG FcRs can be induced in cells infected with human herpes viruses such as herpes simplex virus type 1 (HSV-1)16,17 and human cytomegalovirus (CMV)18-21. Here we demonstrate that FcRs induced by CMV allow immune complexes of HIV to infect fibroblasts otherwise not permissive to HIV infection. Infection was inhibited by prior incubation with human IgG, but not by anti-CD4 antibody or by recombinant soluble CD4. Once HIV had entered CMV-infected cells by means of the FcR, its replication could be enhanced by CMV transactivating factors. Synergism between HIV and herpes viruses could also operate in vivo, enhancing immunosuppression and permitting the spread of HIV to cells not expressing CD4.     

胚胎干细胞及其在人类疾病治疗中的应用

胚胎干细胞是一类多能性干细胞,近年来已成为生命科学研究领域的热点之一．尤其在人类疾病治疗方面有着诱人的应用前景．本文主要介绍了胚胎干细胞在几种疑难疾病治疗上的应用及其前景,胚胎干细胞与异种器官移植。以及目前存在的一些问题．     

Cloned pigs derived from somatic cell nuclear transfer embryos cultured in vitro at low oxygen tension

Great progress have been made in animal cloning in China, as evidenced by the live births of cloned cat- tle[1,2], goats[3,4], and sheep[5]. In contrast, pig cloning is still in its infancy though limited fundamental studieshave been conducted[6]. It is g…     

Dihydroouabain is an antagonist of ouabain inotropic action

The Na+, K+-pump controls a wide variety of cellular systems and its inhibition by cardiac glycosides modifies important physiological functions and evokes several pharmacological effects (refs 1, 2 and refs therein). However, not all the actions of cardiac glycosides can be attributed to Na+, K+-pump inhibition and several observations show that, at low doses, cardiac glycosides stimulate the pump. It has been proposed that their positive inotropic effect could be the sum of two processes: the inhibition of the pump and a still unknown additional inotropic mechanism. In guinea pig heart, low doses of ouabain interact with high-affinity binding sites, which differ from the lower-affinity sites responsible for Na+, K+-pump inhibition. It has been suggested that ouabain interaction with these high-affinity sites could be responsible for the additional inotropic mechanism. The existence of two classes of ouabain-binding sites has been documented not only in guinea pig heart, but also in dog, rat and human heart. Dihydroouabain, a derivative of ouabain in which the lactone ring is saturated, is about 50-fold less potent than ouabain as an inhibitor of Na+, K+-pump and does not stimulate the pump at low doses. Its inotropic effect can be entirely accounted for by the inhibition of the pump. We have examined the pharmacological action of ouabain in the presence of dihydroouabain and report here that dihydroouabain reduces ouabain inotropic action but not Na+, K+-pump inhibition.     

Estimating the human health risk from possible BSE infection of the British sheep flock.

Following the controversial failure of a recent study and the small numbers of animals yet screened for infection, it remains uncertain whether bovine spongiform encephalopathy (BSE) was transmitted to sheep in the past via feed supplements and whether it is still present. Well grounded mathematical and statistical models are therefore essential to integrate the limited and disparate data, to explore uncertainty, and to define data-collection priorities. We analysed the implications of different scenarios of BSE spread in sheep for relative human exposure levels and variant Creutzfeldt-Jakob disease (vCJD) incidence. Here we show that, if BSE entered the sheep population and a degree of transmission occurred, then ongoing public health risks from ovine BSE are likely to be greater than those from cattle, but that any such risk could be reduced by up to 90% through additional restrictions on sheep products entering the food supply. Extending the analysis to consider absolute risk, we estimate the 95% confidence interval for future vCJD mortality to be 50 to 50,000 human deaths considering exposure to bovine BSE alone, with the upper bound increasing to 150,000 once we include exposure from the worst-case ovine BSE scenario examined.     

Production of human lysozyme-transgenic cloned porcine embryos by somatic nuclear transfer

Due to their physiology and organ size, pigs have significant potential as human disease models and as organ transplantation donors. Genetic modification of pigs could provide benefits for both agriculture and human medicine. In this study, five fetal pig fibroblast cell lines from two species (Wuzhishan and Landrace pigs) were transfected using double-marked human lysozyme (HLY) plasmids (pBC1-HLY-GFP-NEO) by a liposome-mediated method. The ratio of green fluorescent protein (GFP)-expressing cells was >95% in sw7, sw8, slw3 and slw6 cell lines, but only 49.3% in slw9 cells. Cells from the four highly transgenic lines were used as nuclear donors to construct embryos, which were then cultured after fusion and activation by electric stimulation. The rate of cleavage was 76.7%, 48 h after activation. After 7 days, 18.5% of cleaved eggs had developed to the blastocyst stage and 93.3% of blastocysts were GFP-positive. These results indicate that transgenic fetal pig fibroblast cell lines could be obtained by a liposome-mediated method, though the transfection efficiency varied between cell lines. Reconstructed embryos derived from transgenic cells could successfully develop into blastocysts, most of which were GFP-positive.     

资本结构与公司价值分析

公司的资本结构在有摩擦的市场环境中，通过削减成本、减少委托代理中的道德风险和诱因问题，改变公司的价值。运用非对称信息博弈方法，分析存在税收和破产成本时，如何进行机制设计，使公司经理通过最大化自己的效用来最大化股东财富，从而确定最佳的负债比率。得出在确定的机制下，好公司较差公司有较高的负债比例和较高的市场价值；合理的资本结构，可以减少公司经理的道德风险，使经理根据股东的意愿行事；高风险的项目能带来更高的公司价值。     

Single amino-acid changes in HIV envelope affect viral tropism and receptor binding

Infection by the human immunodeficiency virus (HIV) is initiated by the binding of its extracellular envelope glycoprotein, gp120, to the CD4 antigen on target cells. To map the residues of the HIV-1 glycoprotein that are critical for binding and to analyse the effects of binding on viral infectivity, we created 15 mutations in a region of gp120 that is important for binding to CD4 (refs 4,5). We find that substitution of a single amino acid (tryptophan at position 432) can abrogate CD4 binding and that virus carrying this mutation is non-infectious. By contrast, other amino-acid changes in the same region do not affect CD4 binding but restrict viral tropism: virions containing isoleucine substitutions at position 425 lose their ability to infect a monocyte cell line (U937 cells) but can still infect T-lymphocyte cell lines (CEM, SUP-T1) and activated human peripheral blood lymphocytes. These results indicate that cellular tropism of HIV can be influenced by a single amino-acid change in gp120.     

Transferrin receptor 1 is a cellular receptor for New World haemorrhagic fever arenaviruses

At least five arenaviruses cause viral haemorrhagic fevers in humans. Lassa virus, an Old World arenavirus, uses the cellular receptor alpha-dystroglycan to infect cells. Machupo, Guanarito, Junin and Sabia viruses are New World haemorrhagic fever viruses that do not use alpha-dystroglycan. Here we show a specific, high-affinity association between transferrin receptor 1 (TfR1) and the entry glycoprotein (GP) of Machupo virus. Expression of human TfR1, but not human transferrin receptor 2, in hamster cell lines markedly enhanced the infection of viruses pseudotyped with the GP of Machupo, Guanarito and Junin viruses, but not with those of Lassa or lymphocytic choriomeningitis viruses. An anti-TfR1 antibody efficiently inhibited the replication of Machupo, Guanarito, Junin and Sabia viruses, but not that of Lassa virus. Iron depletion of culture medium enhanced, and iron supplementation decreased, the efficiency of infection by Junin and Machupo but not Lassa pseudoviruses. These data indicate that TfR1 is a cellular receptor for New World haemorrhagic fever arenaviruses.     

清管器测径板与U型管道中弯头撞击分析

测径清管器经过U型管道时,若测径清管器设计不合理或者清管器运动速度过大,清管器测径板易与弯头相撞,导致测径板变形,从而误以为管道上存在变形。为解决该技术难题,结合一种具体测径清管器,通过分析该测径清管器经过U型管道弯头时的运动特性和测径清管器及弯头的几何特性。得出结论:当测径清管器的运动速度超过一个极限速度时,清管器经过弯头时所受离心力大于清管器支撑板的屈服极限,引起前支撑板屈服,此时测径板可能与弯管内壁外弧侧相撞,同时还得到影响此极限速度值大小的多种因素。另外,发现测径板、支撑板、弯头曲率半径满足一定的几何特性关系时,清管器测径板将与弯管内壁内弧侧撞击。利用得到的这些分析结论,可指导并修正清管器设计,控制测径清管器的运行速度,从而避免测径清管器经过U型管道的弯头时发生撞击。     

混凝沉淀-超滤-消毒组合工艺对二级出水中条件致病菌的去除效能及风险评价

再生水回用是解决水资源危机的重要途径,但污水厂二级出水中存在的大量条件致病菌,会对人体健康构成潜在风险。本研究采用混凝沉淀-UF-NaClO消毒组合工艺作为二级出水的深度处理工艺,探究该组合工艺对条件致病菌（军团菌、铜绿假单胞菌、鸟分支杆菌）和大肠杆菌的去除效能,并以再生水景观回用（划船游湖、孩童玩水、叠水花台观赏）作为评价场景,对条件致病菌进行健康风险评价和参数不确定性因素的敏感性分析。结果表明,混凝沉淀-UF-NaClO消毒组合工艺对水中三种条件致病菌及大肠杆菌的去除效果较好,去除率均能达到98%以上,但出水中的大肠杆菌浓度与三种条件致病菌之间不存在显著相关性,其含量不能够完全地指示三种条件致病菌的含量。二级出水经组合工艺处理后回用于城市景观休闲活动中,三种条件致病菌的单次感染概率较低,对人体健康的安全保障率可达70.9%~100.0%。敏感性分析结果表明：三种暴露途径中感染军团菌和鸟分支杆菌的年健康风险值均受年暴露频率f的不确定性影响最大;划船游湖、孩童玩水两种暴露途径中感染铜绿假单胞菌的年风险值受暴露体积v和铜绿假单胞菌浓度值C的不确定性影响较大。综上,混凝沉淀-UF-NaClO组合工艺可有效控制二级出水中的条件致病菌的浓度,回用于景观休闲活动中的微生物健康风险较低。     

重组痘苗病毒对不同哺乳动物细胞株的感染效率及EGFP表达水平研究

研究重组痘苗病毒对不同哺乳动物细胞的感染效率及表达水平，可为痘苗病毒表达系统宿主细胞的正确选择提供依据．本研究利用重组绿色荧光蛋白基因的痘苗病毒WR—EGFP同时感染不同的哺乳动物细胞株，利用流式细胞仪检测EGFP的表达强度．共使用20种哺乳动物细胞株，其中10种人类组织细胞，2种猴组织细胞。8种小鼠组织细胞．结果表明，重组痘苗病毒wR-EGFP对鼠细胞系BHK21和人细胞系A-549的感染效率和表达效率最佳；整体看，痘苗病毒对多数灵长类动物细胞的感染效率和表达效率优于鼠细胞；对贴壁细胞的感染效率和表达效率明显优于悬浮细胞；但没有特别的组织偏嗜性。