Kinetic measurements of fusion of phosphatidylserine-containing vesicles by electron microscopy and fluorometry

Large unilamellar vesicles (REV) containing phosphatidylserine and phosphatidylethanolamine at a ratio of 1:3 were induced to fuse by adding calcium (4 mM). The kinetics of fusion was monitored by fluorometry using terbium or dipicolinic acid-containing vesicles. The morphology and the states of vesicle aggregation and fusion were examined at approx. 2, 30, 60, 150 and 900 s after calcium addition, by rapid quenching and freeze-fracture electron microscopy. The size and the state of aggregation of vesicles are quantitated from 4000 randomly selected vesicles. The aggregation and fusion kinetics as assayed by fluorescence volume mixing is very well simulated and predicted by the mass action model. The model essentially predicts the time course of the distribution of the aggregates and the increase in size of fused particles as measured by electron microscopy, although in some cases the predicted fusion rate exceeds that by morphometric measurement. No morphological features can be defined as fusion intermediates, although bead-like and rim-like materials may be attributed to the remnants of broken diaphragms between fusion partners.     

血友病患者血清中淋巴腺病病毒/人T细胞Ⅲ型病毒抗体检测

对浙江省1982～1984年注射了美国产浓缩Ⅶ因子制剂的18例血友病患者,用酶联免疫吸附法(ELTSA)检测了血清中淋巴腺病病毒/人T细胞Ⅲ型病毒(LAV/HTLV-Ⅲ)抗体,发现4例阳性,并经免疫荧光试验和Western印迹法证实。2例应用了国产浓缩Ⅷ因子者抗体阴性。一例从美国来华旅游死于艾滋病者,LAV/HTLV-Ⅲ抗体阳性。本研究证明,LAV/HTLVⅢ病毒巳通过美国生产的Ⅷ因子制剂传入中国。     

Action of polyethylene glycol on the fusion of human erythrocyte membranes

Summary Factors affecting the polyethylene glycol (PEG)-induced membrane fusion were examined. Human erythrocyte membrane ghosts, cytoskeleton-free vesicles budded from erythrocytes, mechanically disrupted erythrocyte vesicles, and recombinant vesicles from glycophorin and egg phosphatidylcholine were used as models. Fusion was monitored by darkfield light microscopy and by freeze-fracture electron microscopy. Osmotic swelling was found necessary for fusion between membrane ghosts following PEG treatment. The sample with the highest fusion percentage was sealed ghosts incubated in hypotonic media after at least 5 min of treatment in <25% PEG. At similar osmolarity, glycerol, dextran and PEG produced progressively more pronounced intramembranous particle (IMP) patching, correlating with their increasing fusion percentages. The patching of IMP preceded cell-cell contact, and occurred without direct PEG-protein interaction. The presence of cytoskeletal elements in small vesicles had no significant effect on fusion, nor on the aggregation of intramembranous particle (IMP) upon PEG treatment. Disrupting the membrane by lysolecithin, dimethylsulfoxide, retinol or mild sonication resulted in the fragmentation of ghosts without an increase in fusion percentage. The purity of the commercial PEG used had no apparent effect on fusion. We concluded that the key steps in PEG-induced fusion of cell membrane are the creation of IMP-free zones, and the osmotic swelling of cells after the formation of bilayer contacts during the PEG treatment. Cell cytoskeleton affects PEG-induced fusion only to the extent of affecting IMP patching.     

The biological relevance of HPLC-purified vasoactive intestinal polypeptide monoiodinated at tyrosine 10 or tyrosine 22

Three major forms of monoiodinated VIP (M125I-VIP) were isolated after chloramine-T iodination and HPLC purification. The iodinated tyrosine residue was located in each form of M125I-VIP using arginase C and trypsin digestion for obtaining defined fragments containing only one tyrosine residue. The HPLC isolated iodinated fragments thus obtained were used for HPLC comigration studies with iodinated synthetic C and N terminal VIP fragments and for amino acid analysis. The first two eluting peaks 1 and 2 are (M125I-Tyr10-VIP); peak 1 has an oxidized methionine; peak 3 is a (M125I-Tyr22-VIP) which also has an oxidized methionine. A reduced counterpart of peak 3 named peak 4 was isolated by further HPLC analysis. The ability of the different species of M125I-VIP to stimulate adenosine cyclic 3',5'-phosphate (cAMP) production in transformed colonic cells in culture (HT-29) was compared to that of native VIP. The mean potencies of the M125I-VIP species expressed as a percentage relative to the potency of native VIP were, peak (1): 0.98; (2): 0.84; (3): 1.38; (4): 1.48, in the range of concentrations tested (2-60 pM). The M125I-Tyr22-VIP are significantly more active than native VIP (P less than 0.01). Oxidation of methionine or iodination of tyrosine 10 does not significantly modify the biological activity of VIP. We conclude that iodination of Tyr-22 located in the apolar helical COOH-terminal of VIP increases the effectiveness of VIP interaction with its receptors. Thus the tyrosyl residue and the localized hydrophobic features of VIP are critically involved in the function of this neurotransmitter.     

关于猫颌下腺神经支及其节前神经元的观察

用逆行溃变（Kohnstamm,1902;Yagita,et al.,1909;Torvik,1957）局部电刺激中枢（Chatfield,1942;Magoun et al.,1942;Wang,1943）等方法进行唾液中枢的定位,所得到结果很不一致。近年Satomi（1979）等用辣根过氧化酶（HRP）浸泡猫中间一面神经或鼓索神经,观察了脑干中逆行标记细胞的分布。但用HRP直接浸泡支配猫颌下腺的神经分支尚未见报道。此外,只见到关于鼓索神经纤维类别和数量的分析的光学显微镜研究（Foley,1945）,用光镜和电镜相结合分析颌下腺神经支中的纤     

新疆伊犁五个少数民族的近亲结婚

1983年4月至1984年5月,对新疆伊犁哈萨克自治州五个民族的近亲结婚情况进行了调查。调查的婚姻数为维吾尔2553起,哈萨克1079起,回1235起,锡伯1222起,蒙古446起。近亲结婚率与平均近交系数分别为:维吾尔——8.23％与 46.74×10~(-4),回——8.10％与45.07×10~(-4),锡伯——4.66％与24.93×10~(-4),哈萨克——2.87％与11.31×10~(-4)。蒙古——0.45％与 2.80×10~(-4)。解放以来,维吾尔族与回族的近亲结婚率有上升趋势,而锡伯族的则有下降趋势。各族近亲结婚中大部分为亲表亲结婚,但堂亲结婚在维吾尔族的近亲结婚中竟占26.18％,在回族中也占16.00％。1303名近亲结婚子女中的七岁前死亡率(12.89％)及先天性缺陷与遗传性疾病发生率(2.99％)显著高于6370名非近亲结婚子女的相应值(7.65％及0.38％)。     

琼脂糖平板制备聚焦电泳法进一步提纯胸腺素组分五

用琼脂糖平板等电聚焦电泳法,由胸腺素组分五中分离出三种在聚焦电泳谱上是单一谱线的多肽成份——CP1、CP2和CP3,等电点分别为4.3、4.9和5.6。测定了这些多肽对脐带血中淋巴细胞形成羊红细胞玫瑰花的影响。与对照相比,CP1(2微克/0.6毫升),和CP3(0.2-2微克/毫升)分别在统计学上呈显著和非常显著差异。在相同测定条件下,这三种多肽成份的活性均高于化学合成的胸腺多肽——胸腺素α_1。     

担子菌多糖及羊脾RNA对肿瘤的抑制与有关组织中cAMP含量关系的探讨

本文讨论了对带瘤小鼠注射香菇多糖、松口蘑多糖和羊脾RNA后,活体内有关组织中cAMP含量与抑瘤率之间的关系。 结果表明,上述活性物质不仅增强了机体免疫活性,同时亦促使机体内脾、瘤等组织的cAMP增高,此种增高,与抑瘤率成正相关。     

用双相电泳技术分析棉酚对大鼠成熟精子蛋白质组成的影响

本文首次将双相电泳(等电聚焦和SDS电泳)技术应用于对棉酚作用机理的研究。实验结果表明,棉酚能够改变大鼠成熟精子的蛋白质组成;同时也证明,双相电泳这一技术对棉酚的研究是有效的。     

The groin flap in reparative surgery of the hand

The historical literature of the use of axial vascular pattern flaps from the hypogastric and iliofemoral regions in reparative surgery of the hand is concisely reviewed. Thirty-six iliofemoral (groin) flaps were utilized for delayed primary resurfacing and secondary reconstruction of defects of the hand and forearm. Two flaps (6 percent) were complicated by partial necrosis. We caution against the immediate resurfacing (within 24 hours of injury) of acute crushed hand wounds by distant flaps. The immediate application of a healthy flap on a soiled or crushed wound invites complications of local tissue necrosis, infection, and subsequent loss of the flap. When distant flaps are indicated for coverage of acute hand wounds, delayed primary coverage following complete removal of all nonviable tissue is a safe and reliable regimen. It is advantageous to design the serviceable portion of the flap on the distal area of the vascular territory of the groin flap. Thoughtful yet "radical" defatting can be performed on the lateral portion of the groin flap territory. Constructed in this way, the long medial base of the groin flap allows freedom for movement at the wrist and metacarpophalangeal and interphalangeal joints, thus decreasing edema and stiffness. In the management of soft-tissue defects in the hand requiring distant flap coverage, we choose to utilize the conventional groin flap in preference to the microvascular free flap when both techniques will deliver equal results.