Role of histo-blood group antigens in primate enteric calicivirus infections

Human noroviruses(No V) are associated with large proportion of non-bacterial diarrhea outbreaks together with > 50% of food-associated diarrheas. The function of histo-blood group antigens(HBGAs) in pathogenesis of virus infection was implicated. Until recently however, due to lack of a robust animal and in vitro models of human NoV infection, only the partial knowledge concerning the virus pathogenesis(receptor, coreceptor and target cell) and absence of viable vaccine candidates were the frequently referenced attributes of this acute diarrheal illness. Recently, a novel group of enteric caliciviruses(CV) of rhesus macaque host origin was discovered and described. The new genus within the family Caliciviridae was identified: Rhesus Enteric CV, i.e., "Recovirus"(Re CV). Re CVs are genetically and biologically close relatives of human NoV s, exhibit similar genetic and biological features and are capable of being propagated in cell culture. ReC Vs cause symptomatic disease(diarrhea and fever) in experimentally inoculated macaques. Formulation and evaluation of efficient NoV vaccine might take several years. As suggested by recent studies, inhibition of HBGAs or HBGAbased antivirals could meanwhile be exploited as vaccine alternatives. The purpose of this minireview is to provide the guidance in respect to newly available primate model of enteric CV infection and its similarities with human NoV in utilizing the HBGAs as potential virus co-receptors to indirectly address the unresolved questions of NoV pathogenesis and immunity.

Research Progress on the Large-scale Culture Technology of Mammalian Cells

The culture of mammalian cells is closely related to the development of biotechnology, which has been used extensively in the research and application fields of biology and medical science. In this article, various factors affecting cell cultivation and the application of microcarrier and bioreactor on large-scale culture of mammalian ceils were reviewed.

Humane Non-Human Primate Model of Traumatic Spinal Cord Injury: Quantitative Analysis of Electromyographic Data

A valid non human primate model of traumatic spinal cord injury (TSCI) is essential to evaluate and develop new treatments. In previous experiments, it has been demonstrated that a transmitter can be implanted in the macaque fasicularis monkey that measures electromyographic data from the musculature of the tail. As well, previous experiments have demonstrated that selective lesions can be created in the lower thoracic spinal cord that does not cause limb weakness and/or bowel dysfunction. The histopathological features of these lesions appear similar to human TSCI. This paper describes a method by which the EMG data can be transformed into a quantitative metric of volitional limb movement (“Q”). This metric permits an objective assessment of injury, natural recovery as well as potential efficacy of candidate treatments.

Characteristics of a primate model of focal motor cortical seizures suitable for preclinical testing of therapies like DBS

Background and objective: Generating and characterizing primate models of epileptic seizures are important for understanding pathophysiology of diseases and establishing preclinical efficacy of novel therapies like Deep Brain Stimulation. A focal motor epilepsy model is described here. Method: Seizures were obtained after intracortical penicillin injection into the motor strip through a cannula in two awake monkeys and electrocorticograms were recorded by epidural screws. Seizures were analyzed and compared for number, average duration of each seizure and total duration of ictal activity. Pharmaco-resistance for antiepileptic drug was tested by administration of Diazepam during seizures. Results: A motor status with seizures mimicking those seen in Kojevnikov’s syndrome was easily generated several minutes after penicillin injection and lasted 24 h on an average. The model thus characterized appears stable and consistent. There is no significant variation between experiments in individual primate as well as between two specimens. Diazepam though reduced the total duration of seizures, failed to abolish behavioural seizures. Conclusion: This model represents a good alternative model for preclinical research aiming at testing novel therapies because seizures are obtained on demand, last up to 24 h after a single penicillin injection, are stable and resistant to Diazepam.

2种家养动物朊蛋白基因的扩增与分析

采用PCR方法扩增了云南矮马和北京油鸡的PrP^C基因，并进行序列测定，结合已发表种属朊病毒基因序列，运用分子生物学软件进行同源性分析。结果表明，云南矮马PrP^C基因与已报道的马PrP^C基因比较，其同源性达99％，氨基酸同源性达100％。与牛PrP^C基因比较，同源性达89％，氨基酸同源性达91％。北京油鸡PrP^C基因与鸡的已知序列比较，同源性达99％；与鸭、鸽、鹌鹑的已知序列比较，同源性94％～97％。其翻译的氨基酸序列与鸡的已知序列比较，同源性达99％，与鸭、鸽、鹌鹑比较，氨基酸同源性88％～99％。从本试验结果来看，云南矮马PrP^C基因与牛的PrP^C基因同源性较远，因此感染牛源朊病毒的风险较小，禽类与哺乳动物PrP^C氨基酸属于2个不同的进化分支，因此哺乳动物海绵状脑病不易传染给禽类或引起朊蛋白构型上的改变。这些结果可为海绵状脑病种间屏障补充详细的数据，也为制定相关预防控制措施提供了一些理论依据。

Urokinase-type plasminogen activator is a modulator of synaptic plasticity in the central nervous system:implications for neurorepair in the ischemic brain

The last two decades have witnessed a rapid decrease in mortality due to acute cerebral ischemia that paradoxically has led to a rapid increase in the number of patients that survive an acute ischemic stroke with various degrees of disability.Unfortunately,the lack of an effective therapeutic strategy to promote neurological recovery among stroke survivors has led to a rapidly growing population of disabled patients.Thus,understanding the mechanisms of neurorepair in the ischemic brain is a priority with wide scientific,social and economic implications.Cerebral ischemia has a harmful effect on synaptic structure associated with the development of functional impairment.In agreement with these observations,experimental evidence indicates that synaptic repair underlies the recovery of neurological function following an ischemic stroke.Furthermore,it has become evident that synaptic plasticity is crucial not only during development and learning,but also for synaptic repair after an ischemic insult.The plasminogen activating system is assembled by a cascade of enzymes and their inhibitors initially thought to be solely involved in the generation of plasmin.However,recent work has shown that in the brain this system has an important function regulating the development of synaptic plasticity via mechanisms that not always require plasmin generation.Urokinase-type plasminogen activator(uPA)is a serine proteinase and one of the plasminogen activators,that upon binding to its receptor(uPAR)not only catalyzes the conversion of plasminogen into plasmin on the cell surface,but also activates cell signaling pathways that promote cell migration,proliferation and survival.The role of uPA is the brain is not fully understood.However,it has been reported while uPA and uPAR are abundantly found in the developing central nervous system,in the mature brain their expression is restricted to a limited group of cells.Remarkably,following an ischemic injury to the mature brain the expression of uPA and uPAR increases to levels comparable to those observed during development.More specifically,neurons release uPA during the recovery phase from an ischemic injury,and astrocytes,axonal boutons and dendritic spines recruit uPAR to their plasma membrane.Here we will review recent evidence indicating that binding of uPA to uPAR promotes the repair of synapses damaged by an ischemic injury,with the resultant recovery of neurological function.Furthermore,we will discuss data indicating that treatment with recombinant uPA is a potential therapeutic strategy to promote neurological recovery among ischemic stroke survivors.

Nanog and transcriptional networks in embryonic stem cell pluripotency

象 LIF， BMP 和 Wnt 那样的几个外来的信号能支持胚胎的茎(ES ) 的 theself 更新和 pluripotency 通过调整的房间“ pluripotentgenes。“一个唯一的 homeobox 抄写因素， Nanog，是这些信号的关键下游的受动器之一。Nanog 的提高的水平能维持老鼠 ES 细胞自强独立人士 ofLIF 并且没有喂入器饲料分送器细胞，启用人的 ES 细胞生长。除了外部信号，小径，象 FoxD3 那样的内在的抄写因素， P53 和 Oct4 也涉及调整 Nanog 的表示。机能上地， Nanog 和象 Oct4 和 Sox2 那样的另外的关键 pluripotentfactors 工作控制在 EScell pluripotency 有重要函数的一套目标基因。这些关键因素形成一个规章的网络支持或限制对方“ sexpression 铺平房间，它维持 ES 的性质。

大鼠小脑脑脊液自体输入减少其大脑皮层和海马区的Aβ沉积

目的探讨大鼠小脑脑脊液自体输入对其大脑皮层和海马区β淀粉样蛋白(Aβ)含量的影响及作用机制。方法脑立体定位仪第四脑室抽取脑脊液并注射到同一只大鼠的侧脑室内,饲养1周后,断头法处死大鼠取海马及皮层组织,通过免疫组织化学和酶联免疫吸附试验检测大鼠大脑皮层和海马区Aβ含量,并用Western blot的方法检测胰岛素降解酶(IDE)的表达情况。结果实验组大鼠小脑脑脊液自体输入可以减少自体大脑皮层和海马区Aβ含量,包括可溶性Aβ(P<0.05)和不可溶性Aβ(P<0.05)的含量。同时,IDE的表达较对照组明显增加(P<0.05)。结论大鼠小脑脑脊液自体输入可以降低自体大脑皮层和海马区内的Aβ含量及其造成的病理损伤,这种作用可能是通过上调IDE的表达来实现的。该研究为阿尔茨海默病的临床治疗提供了一个新思路。

E2FBP1 antagonizes the p16^(INK4A)-Rb tumor suppressor machinery for growth suppression and cellular senescence by regulating promyelocytic leukemia protein stability

Cellular senescence is an irreversible cell cycle arrest triggered by the activation of oncogenes or mitogenic signaling as well as the enforced expression of tumor suppressors such as p53, p16INK4A and promyelocytic leukemia protein （PML） in normal cells. E2F-binding protein 1 （E2FBP1）, a transcription regulator for E2F, induces PML reduction and suppresses the formation of PML-nuclear bodies, whereas the down-regulation of E2FBP1 provokes the PML-dependent premature senescence in human normal fibroblasts. Here we report that the depletion of E2FBP1 induces the accumulation of PML through the Ras-dependent activation of MAP kinase signaling. The cellular levels of p16INK4A and p53 are elevated during premature senescence induced by depletion of E2FBP1, and the depletion of p16INK4A, but not p53 rescued senescent cells from growth arrest. Therefore, the premature senescence induced by E2FBP1 depletion is achieved through the pl6INK4A-Rb pathway. Similar to human normal fibroblasts, the growth inhibition induced by E2FBP1 depletion is also observed in human tumor cells with intact p16INK4A and Rb. These results suggest that E2FBP1 functions as a critical antagonist to the pI6INK4A-Rb tumor suppressor machinery by regulating PML stability.

Challenges in animal modelling of mesenchymal stromal cell therapy for inflammatory bowel disease

Utilization of mesenchymal stromal cells(MSCs) for the treatment of Crohn's disease and ulcerative colitis is of translational interest.Safety of MSC therapy has been well demonstrated in early phase clinical trials but efficacy in randomized clinical trials needs to be demonstrated.Understanding MSC mechanisms of action to reduce gut injury and inflammation is necessary to improve current ongoing and future clinical trials.However, two major hurdles impede the direct translation of data derived from animal experiments to the clinical situation:(1) limitations of the currently available animal models of colitis that reflect human inflammatory bowel diseases(IBD).The etiology and progression of human IBD are multifactorial and hence a challenge to mimic in animal models; and(2) Species specific differences in the functionality of MSCs derived from mice versus humans.MSCs derived from mice and humans are not identical in their mechanisms of action in suppressing inflammation.Thus, preclinical animal studies with murine derived MSCs cannot be considered as an exact replica of human MSC based clinical trials.In the present review, we discuss the therapeutic properties of MSCs in preclinical and clinical studies of IBD.We also discuss the challenges and approaches of using appropriate animal models of colitis, not only to study putative MSC therapeutic efficacy and their mechanisms of action, but also the suitability of translating findings derived from such studies to the clinic.

Sodium butyrate prevents radiation-induced cognitive impairment by restoring pCREB/BDNF expression

Sodium butyrate is a histone deacetylase inhibitor that affects various types of brain damages.To investigate the effects of sodium butyrate on hippocampal dysfunction that occurs after whole-brain irradiation in animal models and the effect of sodium butyrate on radiation exposure-induced cognitive impairments,adult C57BL/6 mice were intraperitoneally treated with 0.6 g/kg sodium butyrate before exposure to 10 Gy cranial irradiation.Cognitive impairment in adult C57BL/6 mice was evaluated via an object recognition test 30 days after irradiation.We also detected the expression levels of neurogenic cell markers(doublecortin)and phosphorylated cAMP response element binding protein/brain-derived neurotrophic factor.Radiation-exposed mice had decreased cognitive function and hippocampal doublecortin and phosphorylated cAMP response element binding protein/brain-derived neurotrophic factor expression.Sodium butyrate pretreatment reversed these changes.These findings suggest that sodium butyrate can improve radiation-induced cognitive dysfunction through inhibiting the decrease in hippocampal phosphorylated cAMP response element binding protein/brain-derived neurotrophic factor expression.The study procedures were approved by the Institutional Animal Care and Use Committee of Korea Institute of Radiological Medical Sciences(approval No.KIRAMS16-0002)on December 30,2016.

灵长类中风模型的磁共振成像(英文)

灵长类大脑在结构、生理、功能及神经学上与人类大脑非常相似.因此,灵长类中风模型为研究大脑缺血的损伤机制和开发各种药物治疗方法提供了理想的动物模型.该文介绍了近年来使用的各种灵长类中风模型,并总结了目前可用于该模型的各种磁共振成像(MRI)方法.同时,还讨论了灵长类动物高分辩弥散张量成像(DTI)和磁共振灌注成像(PWI)的数据采集方法和心跳及呼吸引起的生理运动伪影的矫正方法.这些方法改进了弥散一灌注失配(DWI-PWI MISMATCH)成像的精度和质量,有助于提高对脑缺血半暗带演化及其对药物治疗反应的精确评估,为更好地利用灵长类脑中风模型和当代磁共振成像技术进行中风机制研究和药物开发建立一个理想的研究平台.

低密度脂蛋白和高密度脂蛋白相对全蛋黄溶液在猕猴精子低温冷冻中的作用

最近，住各种动物精子低温冷冻的实验中，提取于鸡蛋蛋黄中的低密度脂蛋白被视为优于全蛋黄溶液。与此同时，有研究认为蛋黄中的高密度脂蛋白可能对解冻后精子的生存产生负面影响。关于低密度脂蛋白和高密度脂蛋白在猕猴精子低温冷冻作用的研究还未有过报道。本研究在加和不加渗透型抗冻剂（甘油）两种情况下，通过与全蛋黄溶液对比的手段评价了两种脂蛋白在猕猴精子低温冷冻中的作用。此外，本研究也尝试多种能够改造精于膜与低密度脂蛋白结合体成分的添加剂对保存解冻精子活力的作用。我们的实验结果表明，低密度脂蛋白是伞蛋黄溶液在猕猴精子低温冷冻中发挥保护作用的主要成分。不管有无添加甘油，低密度脂蛋白与令蛋黄溶液在保存解冻精子活性方面作用相近，并未显现出任何优越性。通过加入胆同醇、胆固醇环糊精复合物或卵磷脂来改变精子膜与低密度脂蛋白结合体成分的方法并未改善解冻精子的存活状况；但加入甲基-β-环糊精则明显降低解冻精于的活力。高密度脂蛋白对猕猴精子低温冷冻保存不存在负面影响。对猕猴精子低温冷冻保存而言，本研究表明全蛋黄溶液相比较于稀释液中的有效成分低密度脂蛋白仍然具有优越性。

The plasminogen activating system in the pathogenesis of Alzheimer’s disease

Dementia is a clinical syndrome that affects approximately 47 million people worldwide and is characterized by progressive and irreversible decline of cognitive,behavioral and sesorimotor functions.Alzheimer’s disease(AD)accounts for approximately 60–80%of all cases of dementia,and neuropathologically is characterized by extracellular deposits of insoluble amyloid-β(Aβ)and intracellular aggregates of hyperphosphorylated tau.Significantly,although for a long time it was believed that the extracellular accumulation of Aβwas the culprit of the symptoms observed in these patients,more recent studies have shown that cognitive decline in people suffering this disease is associated with soluble Aβ-induced synaptic dysfunction instead of the formation of insoluble Aβ-containing extracellular plaques.These observations are translationally relevant because soluble Aβ-induced synaptic dysfunction is an early event in AD that precedes neuronal death,and thus is amenable to therapeutic interventions to prevent cognitive decline before the progression to irreversible brain damage.The plasminogen activating(PA)system is an enzymatic cascade that triggers the degradation of fibrin by catalyzing the conversion of plasminogen into plasmin via two serine proteinases:tissue-type plasminogen activator(tPA)and urokinase-type plasminogen activator(uPA).Experimental evidence reported over the last three decades has shown that tPA and uPA play a role in the pathogenesis of AD.However,these studies have focused on the ability of these plasminogen activators to trigger plasmin-induced cleavage of insoluble Aβ-containing extracellular plaques.In contrast,recent evidence indicates that activity-dependent release of uPA from the presynaptic terminal of cerebral cortical neurons protects the synapse from the deleterious effects of soluble Aβvia a mechanism that does not require plasmin generation or the cleavage of Aβfibrils.Below we discuss the role of the PA system in the pathogenesis of AD and the translational relevance of data published to this date.

Urokinase-type plasminogen activator promotes synaptic repair in the ischemic brain

The central nervous system has a very high energy requirement. Accord- ingly, despite representing only 2% of the body＇s mass, the brain uses 20% of the total oxygen consumption. Importantly, because most of this energy is used to maintain synaptic activity, even a mild decrease in its supply to the brain has deleterious implications for synaptic function.

The uPA/uPAR system in astrocytic wound healing

The repair of injured tissue is a highly complex process that involves cell prolife ration,differentiation,and migration.Cell migration requires the dismantling of intercellular contacts in the injured zone and their subsequent reconstitution in the wounded area.Urokinase-type plasminogen activator(u PA)is a serine proteinase found in multiple cell types including endothelial cells,smooth muscle cells,monocytes,and macrophages.A substantial body of experimental evidence with different cell types outside the central nervous system indicates that the binding of uPA to its receptor(uPAR)on the cell surface prompts cell migration by inducing plasmin-mediated degradation of the extracellular matrix.In contrast,although uPA and uPAR are abundantly found in astrocytes and u PA binding to uPAR triggers astrocytic activation,it is unknown if uPA also plays a role in astrocytic migration.Neuronal cadherin is a member of cell adhesion proteins pivotal for the formation of cell-cell conta cts between astrocytes.More specifically,while the extracellular domain of neuronal cadherin interacts with the extracellular domain of neuronal cadherin in neighboring cells,its intracellular domain binds toβ-catenin,which in turn links the complex to the actin cytos keleton.Glycogen synthase kinase 3βis a serine-threonine kinase that prevents the cytoplasmic accumulation ofβ-catenin by inducing its phosphorylation at Ser33,Ser37,and Ser41,thus activating a sequence of events that lead to its proteasomal degradation.The data discussed in this perspective indicate that astrocytes release u PA following a mechanical injury,and that binding of this u PA to uPAR on the cell membrane induces the detachment ofβ-catenin from the intracellular domain of neuronal cadherin by triggering its extracellular signal-regulated kinase 1/2-mediated phosphorylation at Tyr650.Remarkably,this is followed by the cytoplasmic accumulation ofβ-catenin because uPA-induced extracellular signalregulated kinase 1/2 activation also phosphorylates lipoprotein receptor-related protein 6 at Ser1490,which in turn,by recruiting glycogen synthase kinase 3βto its intracellular domain abrogates its effect onβ-catenin.The cytoplasmic accumulation ofβ-catenin is followed by its nuclear translocation,where it induces the expression of uPAR,which is required for the migration of astrocytes from the injured edge into the wounded area.

Outlining key inflammation-associated parameters during early phase of an experimental gram-negative sepsis model in rhesus macaques(Macaca mulatta)

The aim of this study was to identify inflammation-associated markers during the early phase of sepsis in rhesus macaque. Four rhesus macaques were given an intravenous dose of 1010 CFU/kg of E. coli. Blood samples were collected before, or 30 minutes, 2, 4, 6 and 8 hours after E. coli infusion. Physiological parameters, bacteremia, endotoxemia, C-reactive protein(CRP), procalcitonin(PCT), and plasma cytokines/chemokines were determined for each animal. Bacteremia was present in all animals from 30 minutes to 3 hours after E. coli infusion whereas endotoxin was detected during the full-time course. CRP and PCT levels remained at detectable levels during the whole experimental window suggesting an ongoing inflammatory process. Signature cytokines and chemokines such as TNF-α, MIP-1α, and MIP-1β peaked about 2 hours after E. coli infusion and decreased thereafter. Plasma IL-6, IL-12 p40, IFN-γ, and IL-1 Ra, as well as I-TAC, MIG, IP-10 and MCP-1, remained at detectable levels after 4 hours of E. coli infusion. This nonhuman primate model could be useful for the assessment of new therapeutics aiming to suppress key inflammatory markers throughout sepsis early phases.

Middle cerebral artery occlusion methods in rat versus mouse model of transient focal cerebral ischemic stroke

Experimental stroke research commonly employs focal cerebral ischemic rat models （Bederson et al., 1986a; Longa et al., 1989）. In human patients, ischemic stroke typically results from thrombotic or embolic occlusion of a major cerebral artery, usually the mid- dle cerebral artery （MCA）. Experimental focal cerebral ischemia models have been employed to mimic human stroke （Durukan and Tatlisumak, 2007）. Rodent models of focal cerebral ischemia that do not require craniotomy have been developed using intraluminal suture occlusion of the MCA （MCA occlusion, MCAO） （Rosamond et al., 2008）. Furthermore, mouse MCAO models have been wide- ly used and extended to genetic studies of cell death or recovery mechanisms （Liu and McCullough, 2011）. Genetically engineered mouse stroke models are particularly useful for evaluation of isch- emic pathophysiology and the design of new prophylactic, neuro- protective, and therapeutic agents and interventions （Armstead et al., 2010）. During the past two decades, MCAO surgical techniques have been developed that do not reveal surgical techniques for mouse MCAO model engineering. Therefore, we compared MCAO surgical methods in rats and mice.

Mechanistic insights on immunosenescence and chronic immune activation in HIV-tuberculosis co-infection

Immunosenescence is marked by accelerated degradation of host immune responses leading to the onset of opportunistic infections, where senescent T cells show remarkably higher ontogenic defects as compared to healthy T cells. The mechanistic association between T-cell immunosenescence and human immunodeficiency virus(HIV) disease progression, and functional T-cell responses in HIV-tuberculosis(HIV-TB) co-infection remains to be elaborately discussed. Here, we discussed the association of immunosenescence and chronic immune activation in HIV-TB co-infection and reviewed the role played by mediators of immune deterioration in HIV-TB coinfection necessitating the importance of designing therapeutic strategies against HIV disease progression and pathogenesis.

Tissue-type plasminogen activator is a homeostatic regulator of synaptic function in the central nervous system

Membrane depolarization induces the release of the serine proteinase tissue-type plasminogen activator（t PA） from the presynaptic terminal of cerebral cortical neurons.Once in the synaptic cleft this t PA promotes the exocytosis and subsequent endocytic retrieval of glutamate-containing synaptic vesicles,and regulates the postsynaptic response to the presynaptic release of glutamate.Indeed,t PA has a bidirectional effect on the composition of the postsynaptic density（PSD） that does not require plasmin generation or the presynaptic release of glutamate,but varies according to the baseline level of neuronal activity.Hence,in inactive neurons t PA induces phosphorylation and accumulation in the PSD of the Ca^（2＋）/calmodulin-dependent protein kinase IIα（pCa MKIIα）,followed by pCa MKIIα-induced phosphorylation and synaptic recruitment of Glu R1-containing α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid（AMPA） receptors.In contrast,in active neurons with increased levels of pCa MKIIα in the PSD t PA induces pCa MKIIα and p Glu R1 dephosphorylation and their subsequent removal from the PSD.These effects require active synaptic N-methyl-D-aspartate（NMDA） receptors and cyclin-dependent kinase 5（Cdk5）-induced phosphorylation of the protein phosphatase 1（PP1） at T320.These data indicate that t PA is a homeostatic regulator of the postsynaptic response of cerebral cortical neurons to the presynaptic release of glutamate via bidirectional regulation of the pCa MKIIα/PP1 switch in the PSD.