Integrins are not involved in the process of human sperm-oolemmal fusion

BACKGROUND: We investigated whether integrins are required forthe human sperm–oocyte binding and fusion processes. METHODS:The expression of several integrin subunits at the human oocyteplasma membrane was investigated using immunofluorescence microscopy,and the functional role of integrins expressed at the humanoocyte surface in sperm–oocyte interaction was studiedusing a zona-free human oocyte binding and fusion assay. A totalof 144 unfertilized oocytes were stained with anti-integrinantibodies and 147 zona-free unfertilized oocytes were inseminatedin the presence of various anti-integrin antibodies that wereexpressed in oocyte plasma membrane. RESULTS: The antibodiesof six integrin subunits (₂, ₃, ₅, ₆, _V, _M) and six integrinsubunits (₁, ₂, ₃, ₄, ₅, ₆) were bound to the surface of fixedunfertilized oocytes. In contrast, the presence of ₁ and ₄ subunitscould not be verified. The human sperm–oocyte bindingwas only partially inhibited by blocking antibodies of ₂, ₃,₅, ₆, _V, _M, ₁, ₂ and ₃ with a maximum of 55% inhibition, butantibodies of ₄, ₅ and ₆ showed no effect on sperm–oolemmalbinding. A similar reduction of the number of fused sperm wasobserved. However, the ratio of fused sperm to total sperm (boundand fused) was not impaired by all integrin antibodies, suggestingthat integrins had no role in the sperm–oolemmal fusionprocess. CONCLUSIONS: These results suggest that one of thebinding mechanisms can be inhibited by integrin antibodies butthat this mechanism does not play an essential role in the humansperm–oolemmal binding and fusion processes. The othermechanisms, insensitive to integrins, may involve both bindingand fusion processes in human oocytes.     

腰大肌作用力与脊柱伸展应力关系的生物力学实验研究

目的:探讨腰大肌作用力带动脊柱伸展应力的生物力学关系。方法:取家兔12只,分3组,每组4只,解剖后保留枢椎以下完整之脊柱及骨盆、髋关节、上段股骨,不损伤脊柱前、后纵韧带、椎间盘及所附着之腰大肌,保留脊柱背侧的竖脊肌、棘上韧带,置于生物力学拉伸测试仪(日本岛津制作所产AGS-J系列)。上端十字头分别夹枢椎(颈胸腰段)、第1胸椎(胸腰段)和第12胸椎(腰段),下端十字头夹股骨上部;分别作有腰大肌状态下和切断腰大肌状态下,股髋自屈曲位到过伸带动脊柱自屈曲位到过伸位拉伸试验,测定两种不同状态下脊柱各节段的伸展应力(N/mm2)。结果:有腰大肌状态和切断腰大肌状态下,股-髋-脊柱拉伸后脊柱伸展应力分别为:颈胸腰全段平均为306.6675N/mm2:78.7167N/mm2;胸腰段为680.8417N/mm2:373.0375N/mm2;腰段为1990.7944N/mm2:523.0608N/mm2;经统计学分析,具显著性差异,P<0.01。结论:腰大肌作用力对脊柱伸展应力影响显著,颈胸腰段占74.33%、胸腰段占45.21%,腰段占73.73%的伸展应力源自腰大肌。脊柱在腰大肌作用下产生腰椎向腹部的弯曲。     

Action of remantadine on fusion of the lipid envelope of influenza a virus with plasma and internal membranes in lymphoblastoid cells

Department of Biomembranes, Research Center for Development and Introduction of Modern Methods of Molecular Diagnosis, Ministry of Health of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR I. P. Ashmarin.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 109, No. 5, pp. 483–485, May, 1990.     

人KIR2DL1-Ig融合蛋白在COS-7细胞的表达

目的：获得人KIR2DL1分子(killer lg—like receptor 2DL1)胞外区与人IgG Fc段的融合蛋白。方法：从人外周血单个核细胞中提取总RNA，通过RT—PCR扩增编码KIR2DL1胞外段cDNA，经Nhe Ⅰ和BamH Ⅰ双酶切后，定向插入真核细胞表达载体CD51negl中。构建的重组真核表达载体CD51negl—KIR2DL1。经酶切分析和测序鉴定后，通过DEAE—dextran／ehloroquine法转染COS—7细胞：瞬时表达后，取培养上清液经亲和层析、ELISA、SDS—PAGE及Western印迹，鉴定融合蛋白的表达及其免疫学活性。结果：序列测定证实，该重组表达载体含有正确的KIR2DL1胞外区基因序列。重组真核表达载体CD5lnegl—KIR2DL1转染COS—7细胞后，ELISA法检测细胞培养上清中有融合蛋白的表达。SDS—PAGE结果显示，该融合蛋白的相对分子质量(Mc)约为73000。Western印迹结果证实，该蛋白能被特异性单克隆抗体(mAb)EB6所识别。结论：KIR2DL1—Ig融合蛋白表达载体成功构建并在COS—7细胞中获得功能性表达，为KIR2DL1的功能及其配体MHC的研究奠定了基础。     

手术治疗46例胸腰椎爆裂骨折的临床分析

目的探讨前路、后路、前后路联合手术治疗胸腰椎爆裂骨折的特点。方法依据对患者所选择的前路、后路、前后路联合手术术式分为三组,进行影像学评价和神经功能评价。结果各组末次随访脊髓功能评价分级提高情况采用R×C表x2检查方法进行统计学分析,各组差异无显著意义。各组术后cobb氏角改善率应用秩和检验方法进行统计学分析,各组差异无显著意义。结论我们综合应用Denis和McAfee的分型,结合了骨折形态、损伤机制和稳定性评价,对胸腰椎爆裂骨折的治疗有较好的指导意义。手术方式的选择更多基于脊柱机械性稳定性、神经性稳定性评价。综合考虑骨折部位、骨折后时间、患者年龄、工种以及术者对入路的熟悉程度等。     

不同位置植入SMH人工腰椎间盘对小关节应力分布的影响

目的:通过三维有限元方法研究正常椎间盘、SMH人工腰椎间盘前置、中置和后置四组模型中小关节的应力分布情况,探讨不同位置植入SMH人工腰椎间盘对小关节应力分布的影响。方法:建立正常腰椎间盘、人工SMH腰椎间盘前置、中置和后置的三维模型,然后模拟腰椎节段的运动,进行小关节应力分布的比较研究。结果:与正常L4/5椎间盘模型相比,除SMH人工腰椎间盘前置模型前屈时比正常模型小关节应力小外,其余运动时SMH人工腰椎间盘前置、中置和后置模型中小关节的应力均高于正常腰椎间盘组,但后置SMH人工腰椎间盘组则相对较小。结论:SMH人工腰椎间盘后置可以更好地预防小关节的退变;人工椎间盘植入位置的不同对人工腰椎间盘术远期疗效的预测具一定的作用。     

前路椎体次全切在治疗颈椎后纵韧带骨化中的应用

目的：探讨颈前路椎体次全切治疗颈椎后纵韧带骨化的手术减压范围。方法：采用前路椎体次全切植骨融合术治疗颈椎后纵韧带骨化56例，其中完全切除骨化者47例，用“漂浮法”处理者9例，并针对不同个体及病变特点采用不同的减压范围。结果：54例获得3个月-6a随访，平均28个月。植骨均于术后3-5个月内获得骨性融合。JOA评分由术前8．5分提高到术后14．1，平均改善率74％，优良率80．2％。结论：行椎体次全切术治疗颈椎后纵韧带骨化时应针对不同个体及病变特点采用不同的足够的减压范围，可以减少并发症，并获得较佳的疗效。     

Conformational changes in Sindbis virions resulting from exposure to low pH and interactions with cells suggest that cell penetration may occur at the cell surface in the absence of membrane fusion

Alphaviruses have the ability to induce cell-cell fusion after exposure to acid pH. This observation has served as an article of proof that these membrane-containing viruses infect cells by fusion of the virus membrane with a host cell membrane upon exposure to acid pH after incorporation into a cell endosome. We have investigated the requirements for the induction of virus-mediated, low pH-induced cell-cell fusion and cell-virus fusion. We have correlated the pH requirements for this process to structural changes they produce in the virus by electron cryo-microscopy. We found that exposure to acid pH was required to establish conditions for membrane fusion but that membrane fusion did not occur until return to neutral pH. Electron cryo-microscopy revealed dramatic changes in the structure of the virion as it was moved to acid pH and then returned to neutral pH. None of these treatments resulted in the disassembly of the virus protein icosahedral shell that is a requisite for the process of virus membrane-cell membrane fusion. The appearance of a prominent protruding structure upon exposure to acid pH and its disappearance upon return to neutral pH suggested that the production of a "pore"-like structure at the fivefold axis may facilitate cell penetration as has been proposed for polio (J. Virol. 74 (2000) 1342) and human rhino virus (Mol. Cell 10 (2002) 317). This transient structural change also provided an explanation for how membrane fusion occurs after return to neutral pH. Examination of virus-cell complexes at neutral pH supported the contention that infection occurs at the cell surface at neutral pH by the production of a virus structure that breaches the plasma membrane bilayer. These data suggest an alternative route of infection for Sindbis virus that occurs by a process that does not involve membrane fusion and does not require disassembly of the virus protein shell.     

Cytogenetics of a new cytotype of African Mus (subgenus Nannomys) minutoides (Rodentia, Muridae) from Kenya: C- and G- banding and distribution of (TTAGGG) n telomeric sequences

We present the results of a cytogenetic study on Mus (Nannomys) minutoides from Kenya by means of C- and G- banding and in-situ fluorescence hybridization (FISH) to localize the telomeric sequences. The karyotype is characterized by the occurrence of several Rb chromosomes Rb(1.X), Rb(1.Y). Rb(2.17), Rb(3.13), Rb(4.10), Rb(5.11), Rb(6.7), Rb(8.12), not previously described for this species. This finding suggests a high level of chromosomal diversification, which means it is possible to consider this cytotype as a new, well-differentiated, chromosomal lineage within the subgenus. The C-banding of the metaphases illustrated conspicuous blocks of centromeric heterochromatin at the paracentromeric regions of all telocentric chromosomes. Centromeric heterochromatin is not visible on all biarmed chromosomes. Following hybridization with telomeric probes, bright interstitial telomeric sequence (ITS) fluorescence signals are evident at the pericentromeric area of all Rb chromosomes, with the exception of Rb(2.17). Considering the localization of the C-positive heterochromatin and of the telomeric sequences, the events leading to the Kenyan cytotype from an all-telocentric condition probably included two steps: first, fusion without loss of heterochromatin and pericentromeric telomeric sequences; second, the reduction of the C-positive satellite DNA followed by the amplification of telomeric sequences in the C-negative paracentromeric region of Rb chromosomes. The presence of a single Rb(2.17) without ITS indicates possible variations of this mechanism.