人心连续组织切片的计算机三维重建

用表面重建法对人心连续组织切片进行了三维重建，重建中应用了火棉胶断面切片技术及图像拟合与插值处理，与以往的解剖断面重建比较，图像更加清晰、逼真，可从不同观察心脏的外部轮廓与内部结构。作为三维重建的基础性研究，可为心血管影像诊断提供解剖形态学依据和可靠的三维软件。     

鸡卵黄抗体的制备以及金磁微粒为载体去除人血浆部分高丰度蛋白的研究

目的:以人血浆白蛋白(HSA)和IgG为免疫原,制备特异性鸡卵黄抗体IgY(Egg yolk immunoglobulin),并将其固定于金磁微粒表面,用于HSA和IgG的去除研究.方法:用HSA、IgG以及混合成分分别作免疫原免疫Roman母鸡.制备抗HSA和IgG鸡卵黄抗体IgY,并对IgY的分离提取条件进行优化.SDS-PAGE和间接ELISA检测IgY的纯度和效价.将高效价IgY固定于金磁颗粒表面进行血浆高丰度蛋白去除研究.结果:免疫后60～120 d内,鸡血清抗体效价可达1∶15 000～1∶25 000;收获鸡蛋,提取得到的卵黄抗体IgY效价可达1∶10000～1∶25000,纯度98%以上;采用金磁微粒载体固定IgY,可对血浆中的HSA,IgG进行特异性的去除.结论:人血浆中的高丰度蛋白成分HSA和IgG免疫产蛋母鸡后,可从鸡卵黄中分离提取到高效价、高纯度的卵黄抗体IgY;IgY偶联于金磁微粒表面可特异性的去除人血浆中的HSA和IgG,作为血浆蛋白质组学研究的一种新方法,有较好的应用前景.     

小胶质细胞极化介导炎症反应在脊髓损伤中的作用

背景:小胶质细胞极化参与脊髓损伤后的炎症反应,并在其中发挥关键作用。相关研究表明,有效诱导小胶质细胞从M1促炎表型向M2抗炎表型极化,可以减轻脊髓损伤后的炎症反应,促进组织的修复再生和神经功能的恢复。目的:文章对小胶质细胞的功能和极化、小胶质细胞极化对脊髓损伤的影响及其潜在调控策略以及脊髓损伤后炎症反应进行综述。方法:检索PubMed、Web of Science和中国知网数据库,英文检索词为“microglia,polarization,spinal cord injury,inflammation”,中文检索词为“小胶质细胞、极化、脊髓损伤、炎症”,按纳入和排除标准共纳入80篇文献进行总结。结果与结论:①由小胶质细胞介导的稳定而持续的炎症反应,对脊髓损伤的预后至关重要。②在生理条件下,小胶质细胞处于M0静止表型,但在脊髓损伤后,小胶质细胞活化,进而极化成M1促炎表型,导致神经组织修复能力降低和出现持续性神经炎症。③在脊髓损伤的炎症反应过程中,调控小胶质细胞向M2表型极化或至少向M2表型倾斜,有利于抑制氧化应激反应、调节突触重塑、促进轴突再生和血管生成,是一种有效的调控策略。④截止到目前的研究表明,间充质干细胞、外泌体、临床药物、天然产物、miRNAs和靶点分子可调控小胶质细胞在M1和M2表型之间的转换,这为脊髓损伤后神经组织的修复提供了一种新的思路,未来需进一步研究小胶质细胞在脊髓损伤过程中调控极化的详细机制。     

眼底图像畸变的校正及多幅图像的特征点拼接算法

对于眼底图像 ,由于不在正视点取像造成的几何畸变和由于荧光素从血管壁渗漏等造成的灰度畸变 ,几乎是不可避免的。为临床诊断提供丰富信息的需要出发 ,进行多幅眼底图像的拼接是必要的。本文为此提出了几个新的处理方法 ,如基于大窗口的滤波灰度校正方法、基于透视投影原理的几何校正方法与基于小波子空间上的拼接算法 ,均具有效果好和耗时少的优点。     

个性化截骨工具辅助下人工全膝关节置换与传统人工全膝关节置换手术精确度及临床疗效比较

目的比较3D打印个性化截骨工具辅助(patient-specific instrumentation,PSI)下人工全膝关节置换术(total knee arthroplasty,TKA)与传统TKA的手术精确度和临床疗效。方法自2017年9月至2018年12月,将40例拟接受初次膝关节置换患者随机分为2组,每组均为20人。一组应用个性化截骨工具辅助TKA手术(PSI组),另一组接受传统TKA(对照组)。比较两组患者的冠状面下肢机械轴线、手术时间、术中出血量、术后引流量以及HSS评分。结果对照组和PSI组的手术时间分别为(103.3±18.7) min和(91.3±15.7) min;术中出血量分别为(372.0±53.0)mL和(332.8±47.0)mL;术后引流量分别为(378.8±97.2)mL和(315.0±89.0)mL。两组手术时间、术中出血量、术后引流量比较差异均有统计学意义(P0.05)。对照组和PSI组术后2周HSS评分分别为(89.3±2.8)分和(88.7±2.9)分,两组比较差异无统计学意义(P0.05)。对照组和PSI组术后全下肢力线差值分别为(1.9±1.1)°和(1.2±1.0)°,冠状面股骨假体角度(frontal femoral component angle,FFC)差值分别为(2.1±1.1)°和(1.1±0.9)°,两组比较差异有统计学意义(P0.05)。冠状面胫骨假体角度(frontal tibia component angle,FTC)差值分别为(1.3±0.8)°和(1.4±0.8)°,两组比较差异无统计学意义(P0.05)。结论PSI辅助TKA较传统TKA手术时间更短、术中出血量更少,并且术后冠状面全下肢力线及股骨力线的改善优于传统TKA手术。     

High density of immobilized galactose ligand enhances hepatocyte attachment and function

Galactosylated surface is an attractive substrate for hepatocyte culture because of the specific interaction between the galactose ligand and the asialoglycoprotein receptor on hepatocytes. In this study, we described a scheme to achieve high density of immobilized galactose ligands on polyethylene terephthalate (PET) surface by first surface-grafting polyacrylic acid on plasma-pretreated PET film under UV irradiation, followed by conjugation of a galactose derivative (1-O-(6'-aminohexyl)-D-galactopyranoside) to the grafted polyacrylic acid chains. A high galactose density of 513 nmol/cm(2) on the PET surface was used in this study to investigate the behavior of cultured hepatocyte. This engineered substrate showed high affinity to fluorescein isothiocyanate-lectin binding. Primary rat hepatocytes, when seeded at a density of 2 x 10(5) cells/cm(2), attached to the galactosylated PET substrate at a similar efficiency compared with collagen-coated substrate. The hepatocytes spontaneously formed aggregates 1 day after cell seeding and showed better maintenance of albumin secretion and urea synthesis functions than those cultured on collagen-coated surface.     

The leptospiral major outer membrane protein LipL32 is a lipoprotein expressed during mammalian infection

We report the cloning of the gene encoding the 32-kDa lipoprotein, designated LipL32, the most prominent protein in the leptospiral protein profile. We obtained the N-terminal amino acid sequence of a staphylococcal V8 proteolytic-digest fragment to design an oligonucleotide probe. A Lambda-Zap II library containing EcoRI fragments of Leptospira kirschneri DNA was screened, and a 5.0-kb DNA fragment which contained the entire structural lipL32 gene was identified. Several lines of evidence indicate that LipL32 is lipid modified in a manner similar to that of other procaryotic lipoproteins. The deduced amino acid sequence of LipL32 would encode a 272-amino-acid polypeptide with a 19-amino-acid signal peptide, followed by a lipoprotein signal peptidase cleavage site. LipL32 is intrinsically labeled during incubation of L. kirschneri in media containing [(3)H]palmitate. The linkage of palmitate and the amino-terminal cysteine of LipL32 is acid labile. LipL32 is completely solubilized by Triton X-114 extraction of L. kirschneri; phase separation results in partitioning of LipL32 exclusively into the hydrophobic, detergent phase, indicating that it is a component of the leptospiral outer membrane. CaCl(2) (20 mM) must be present during phase separation for recovery of LipL32. LipL32 is expressed not only during cultivation but also during mammalian infection. Immunohistochemistry demonstrated intense LipL32 reactivity with L. kirschneri infecting proximal tubules of hamster kidneys. LipL32 is also a prominent immunogen during human leptospirosis. The sequence and expression of LipL32 is highly conserved among pathogenic Leptospira species. These findings indicate that LipL32 may be important in the pathogenesis, diagnosis, and prevention of leptospirosis.     

粉防己碱预处理对心肌缺血/再灌注损伤大鼠肿瘤坏死因子-α表达及核因子活化的影响

目的探讨粉防己碱预处理对心肌缺血/再灌注过程中大鼠肿瘤坏死因子-α表达及与核因子活化的影响以及相关关系。方法60只SD大鼠随机分为3组:缺血/再灌注损伤组、假手术组、粉防己碱治疗组,缺血/再灌注损伤组结扎大鼠左前降支造成心肌缺血30min后再灌注24h后处死大鼠;假手术组只穿针不结扎,余步骤同缺血/再灌注损伤组;粉防己碱治疗组在缺血前30min腹腔注射粉防己碱,余步骤同缺血/再灌注损伤组。24h后取心肌标本,用酶联免疫吸附法检测心肌组织中肿瘤坏死因子-α表达水平,凝胶电泳迁移率变化分析检测核因子的活性。结果粉防己碱组与缺血再灌注组相比肿瘤坏死因子-α表达水平明显减低(208.40±25.12 VS 306.65±17.78,P<0.01)而与假手术组相比表达明显增强(208.40±25.12 VS 61.45±9.20,P<0.01)。粉防己碱组核因子的活性明显低于缺血再灌注组(29.58±1.56 VS 40.33±4.39,P<0.01)而与假手术组无显著性差异(29.58±1.56 VS 30.09±3.46,P>0.05)。结论粉防己碱预处理可以使心肌缺血/再灌注损伤过程中肿瘤坏死因子-α生成明显减少,核因子的活化受到有效抑制,从而起到减轻心肌缺血/再灌注损伤的作用。     

Umbilical cord blood transplantation in adults using myeloablative and nonmyeloablative preparative regimens.

Unrelated umbilical cord blood (UCB) transplantation has recently been explored in an increasing number of adult patients. The relative ease of procurement and the lower-than-anticipated risk of severe acute graft-versus-host disease has made UCB transplantation an appealing alternative to bone marrow-derived hematopoietic stem cells. The use of reduced-intensity or nonmyeloablative preparative regimens to allow engraftment of UCB broadens the scope of patients who may benefit from allogeneic immunotherapy, including elderly and medically infirm patients with no matched sibling donor. This review summarizes the available data on the use of UCB as an alternative source of hematopoietic stem cell transplantation in adult patients.     

Characterization of virus-like particles assembled in a recombinant baculovirus system expressing the capsid protein of a fish nodavirus

Infectious bursal disease virus (IBDV) causes severe immunodeficiency in young chickens by destroying the precursors of antibody-producing B cells in the bursa of Fabricius. It has been shown that IBDV infection induces apoptosis in chicken embryo and tissue culture cells. We previously reported that an IBDV mutant lacking the expression of 17-kDa nonstructural (NS) protein exhibited decreased apoptotic effects in cell culture as compared to the parental IBDV, suggesting that the NS protein may be involved in induction of apoptosis. Here, we report that the NS protein of IBDV alone is capable of inducing apoptosis in cell culture. Transfection of chicken B-lymphocyte cell line (RP9) and chicken embryo fibroblast cells with a plasmid DNA, containing the NS protein gene under the control of the immediate-early promoter-enhancer region of human cytomegalovirus, induced programmed cell death in both cell lines. Apoptosis changes, such as chromatin condensation, DNA fragmentation, and the appearance of apoptotic nuclear bodies, were observed in cell cultures 48-h posttransfection. As reported earlier, the mutant IBDV grew to lower titers with slower replication kinetics and lower cytopathogenicity when compared to that of the parental virus. Here, we demonstrate that the mutant virus is closely associated with cells and its yield from the supernatant was approximately 30-fold lower than the wild-type due to increased cell association, indicating a deficiency in lysis of virus-infected cells. Taken together, our results indicate that the NS protein of IBDV is highly cytotoxic, which brings about the release of the viral progeny from cells, and thus play an important role in viral pathogenesis.