细粒棘球绦虫精子形成的超微结构观察

用透射电镜观察了细粒棘球绦虫精原细胞、精母细胞、精细胞的发育变化和精子的主要形态结构特点，从而揭示了该虫精子的形成过程。结果显示细粒棘球绦虫的精子形成主要是由于精细胞内微管结构的多次排列组合形成的。精细胞和精子中无线粒体和顶体结构。成熟的精子可分为头部和尾部。头部色较淡，顶端有与精子轴丝呈30°角排列的明暗相间的带构成的帽状物包裹。完整精子结构为固有膜包统一条轴丝，轴丝向前伸入头部，向后延伸至尾部。固有膜内线有微管排列，尾部前段一例可有2层做管，多为“2×4＋1，”结构，后段为一层微管紧贴固有腹内缘。抽丝有一个中央微管和9对外周微管组成，中央微管的外围有一个纤维鞘包绕，由纤维鞘发出纤维丝与外周9对微管相连，形成一个车轮状的“9＋1”微管系统结构。     

带绦虫精子发生过程中精子变形的超微结构观察

目的：研究带绦虫在精子发生过程中的精子变形。方法：透射电镜。结果：32个玫瑰花样的次级精母细胞经第2次减数分裂后,产生64个精细原形体（64－spermatid－plasmodium）。精细胞的变形过程复杂,首先见精细胞胞质和核伸长,胞质增多由胞质桥“cytophore”相连;然后是核的进一步伸长,核染色质聚合成线束状,最后脱离胞质桥,形成成熟精子。成熟的精子呈细线状,长约16．2－18．6μm,宽0．35－0．45μm,可分为有核的头部及无核的尾部两部分。头部长约5－6μm,占体长的1／3,有一个较长的核缠绕着轴丝,无线粒体。尾部长约11．2－16．6μm。在尾部的前段及头部的后方,纵断面上见一些线粒体包绕着轴丝,全长约1．6－1．7μm。精子的尾部只含一条结构为“9＋1”的轴丝。精子横断面,质膜下见46条微管（周围微管）。结论：带绦虫精子子的变形是一个非常复杂的过程。     

日本血吸虫精子的超微结构

目的：了解日本血吸虫正常精子的超微结构。方法：半超薄切片定位雄虫睾丸和含有成熟精子的雌虫输卵管，常规透射电镜制样并观察。结果：日本血吸虫精子由头、尾两部分组成，头部呈长卵圆形，平均长6．2μm，宽1．4μm，无顶体构造，前端钝圆，后端尖细，质膜下有1圈纵行的微管，核1个，前端有少量线粒体，尾部鞭毛1根，在中、后段主体部分鞭毛轴丝外周为9组二联管，中央为一团弥散的电子致密物质，呈9×2＋《1》型；但在过渡区鞭毛轴丝中央无电子致密物质，呈9×2＋0型。结论：日本血吸虫精子超微结构与其他血吸虫相似，具有同源性，但明显区别于复殖目大多数吸虫的构造。     

云南斯氏狸殖吸虫染色体核型的研究

目的了解云南斯氏狸殖吸虫染色体核型。方法取云南斯氏狸殖吸虫成虫卵巢和睾丸,经短期细胞培养后制作染色体标本片,显微镜观察并照相。结果云南斯氏狸殖吸虫的染色体核型为n=11和2n=22。结论云南斯氏狸殖吸虫染色体核型与四川斯氏狸殖吸虫相同。     

从形态变化,生活史和DNA检测排除泡囊狸殖的独立性研究

目的通过形态观察、生活史循环试验和分子生物学鉴定研究泡囊狸殖吸虫的独立性问题。方法从三元区采集溪蟹分离得到泡囊狸殖吸虫囊蚴和斯氏狸殖吸虫囊蚴,进行形态观察。以泡囊狸殖吸虫虫卵投放到调查过无泡囊狸殖吸虫的疫区中感染螺蛳,继而感染溪蟹,稍后检查蟹体内所获囊蚴是否保持泡囊狸殖吸虫形态还是同斯氏狸殖吸虫囊蚴近似。提取斯氏狸殖吸虫囊蚴和泡囊狸殖吸虫囊蚴的基因组DNA,用特异引物,PCR扩增其ITS2基因并将PCR扩增产物纯化后测序,然后通过GenBank的Blast程序分析基因序列的同源性,并应用MEGA3.0软件中的ME程序构建种系发生树。结果泡囊狸殖吸虫囊蚴与斯氏狸殖吸虫囊蚴形态差异明显,但观察到泡囊狸殖吸虫囊蚴在从蟹体分离后0.5~2 h内有57.14%变成同斯氏狸殖吸虫囊蚴近似的特征。将泡囊狸殖吸虫虫卵投放到自然环境进行生活史循环试验,从收集的溪蟹中只分离到斯氏狸殖吸虫囊蚴而没有发现泡囊狸殖吸虫囊蚴。基因同源性比较结果显示:泡囊狸殖吸虫与斯氏狸殖吸虫的ITS2基因同源性为100%,碱基差异为0,没有基因差异。结论虽然泡囊狸殖吸虫囊蚴和斯氏狸殖吸虫囊蚴的形态特征有较明显的差别,但放置一段时间后,57.14%的泡囊狸殖吸虫囊蚴变成与斯氏狸殖吸虫囊蚴形态相似的特征。从用泡囊狸殖吸虫虫卵进行生活史循环试验中收集的溪蟹体内没有发现泡囊狸殖吸虫囊蚴。ITS2基因序列的同源性比较分析发现斯氏狸殖吸虫和泡囊狸殖吸虫的基因高度同源,而且种系发生树也显示二者之间无明显的遗传分化。因此,斯氏狸殖吸虫和泡囊狸殖吸虫为同一物种,泡囊狸殖吸虫不具有虫种独立性。     

斯氏狸殖吸虫转换宿主的研究

斯氏狸殖吸虫 195 9年由陈心陶首次报道 ,是中国独有虫种。我们在狸殖吸虫宿主转换的系列研究中 ,对犬与斯氏狸殖吸虫的生物学关系及宿主相容性进行了研究 ,本文报道斯氏狸殖吸虫在犬体内的分布、发育与转换宿主的特点。1　材料与方法从湖北省十堰市采集锯齿华溪蟹 (Sinopotamondenticu latum)中分离斯氏狸殖吸虫囊蚴。囊蚴感染豚鼠 5 0d后 ,用生理盐水逸出法收集各部位的童虫 ,在林格氏液内保存备用。将发育较小和发育较大的童虫分别感染家犬 1只 ,6 0d后剖杀 ,收集虫体观察。2　结　　果斯氏狸殖吸虫在鼠与犬间的宿主转换囊蚴感染豚鼠5 0…     

斯氏狸殖吸虫抗原分析和血清学诊断方法的研究

[目的 ]分析斯氏狸殖吸虫囊蚴、童虫和成虫各期抗原和建立特异性抗原的斯氏狸殖吸虫病血清学诊断方法。 [方法 ]用SDS PAGE分离斯氏狸殖吸虫的各虫期抗原 ,经免疫印迹识别成虫期特异性诊断抗原。采用电洗脱技术分离 10～ 30kDa蛋白组分 ,建立纯化抗原的dot ELISA血清学诊断斯氏狸殖吸虫病。 [结果 ]斯氏狸殖吸虫病患者血清与成虫抗原的 10～ 30kDa显示较多免疫识别带 ,主带为 2 2、2 4和 2 6kDa。与血吸虫病和华支睾吸虫病患者血清的交叉反应带出现于 6 0～ 90kDa。斯氏狸殖吸虫成虫 10～ 30kDa抗原的dot ELISA与成虫粗抗原的ELISA检测 2 8例疑诊病人血清 ,两法阳性率间差异无显著性。而检测 38例感染其它吸虫和肺部疾病患者血清 ,粗抗原的ELISA交叉反应率为 13 2 % (5 /38)。 [结论 ]斯氏狸殖吸虫成虫 10～ 30kDa抗原的dot ELISA为斯氏狸殖吸虫病高度特异和敏感的血清学诊断方法。     

斯氏狸殖吸虫成虫可溶性抗原蛋白质分析

目的分析和检测斯氏狸殖吸虫成虫可溶性抗原的特异抗原组分。为建立特异、敏感的免疫学诊断方法、研制抗斯氏狸殖吸虫药物和疫苗提供理论基础。方法用聚丙烯酰胺凝胶电泳（SDS—PAGE）、双向电泳（Two—dimensional gel eleetrophoresis，2-DE）和免疫印迹（Western blot）等方法，对斯氏狸殖吸虫成虫可溶性抗原进行分析。结果SDS-PAGE分离出22条蛋白带，其中主带8条，分子质量单位为13～64ku，未见65ku以上条带。Western blot显示20条显色带，主带6条，分子质量单位分别为13、18、22、28、35和64ku。2-DE电泳分离出斯氏狸殖吸虫成虫可溶性抗原48个多肽斑点，分子质量单位为14～70ku，绝大部分分布于pI3．10～7．14，少数分布于pI8．78～9．85，在pI7．14～8．78之间几乎未见多肽斑点。Western blot分析出其中的10个主要多肽斑点能被斯氏狸殖吸虫病患者血清识别，大部分为集中在酸性区域的小分子多肽。结论斯氏狸殖吸虫成虫可溶性抗原经SDS-PAGE电泳分离出的22条蛋白带中，有6条蛋白含量较高，能被斯氏狸殖吸虫病患者血清识别。2-DE电泳分离出的48个多肽斑点中，有10个含量较高，能被斯氏狸殖吸虫病患者血清识别，大部分为偏酸性的小分子多肽。     

云南省大理州斯氏狸殖吸虫病例报告

目的报道斯氏狸殖吸虫病5例。方法描述分析斯氏狸殖吸虫病5例患者的流行病学、临床表现、实验室检查、诊治及愈后。结果5病例均有食生螃蟹史,肺吸虫抗体（IgG）均阳性,1例检出斯氏狸殖吸虫幼虫1条（2×2．8mm）。采用吡喹酮、丙硫咪唑治疗斯氏狸殖吸虫病疗效显著,预后良好。结论斯氏狸殖吸虫病临床表现复杂多样,极易造成误诊,流行区应注意及时诊治     

斯氏狸殖吸虫成虫可溶性抗原蛋白质分析

目的分析和检测斯氏狸殖吸虫成虫可溶性抗原的特异抗原组分,为建立特异、敏感的免疫学诊断方法、研制抗斯氏狸殖吸虫药物和疫苗提供理论基础。方法用聚丙烯酰胺凝胶电泳(SDS-PAGE)、双向电泳(Two-dimension-al gel electrophoresis,2-DE)和免疫印迹(Western blot)等方法,对斯氏狸殖吸虫成虫可溶性抗原进行分析。结果SDS-PAGE分离出22条蛋白带,其中主带8条,分子质量单位为13~64 ku,未见65 ku以上条带。Western blot显示20条显色带,主带6条,分子质量单位分别为13、18、22、28、35和64 ku。2-DE电泳分离出斯氏狸殖吸虫成虫可溶性抗原48个多肽斑点,分子质量单位为14~70 ku,绝大部分分布于pI 3.10~7.14,少数分布于pI 8.78~9.85,在pI 7.14~8.78之间几乎未见多肽斑点。Western blot分析出其中的10个主要多肽斑点能被斯氏狸殖吸虫病患者血清识别,大部分为集中在酸性区域的小分子多肽。结论斯氏狸殖吸虫成虫可溶性抗原经SDS-PAGE电泳分离出的22条蛋白带中,有6条蛋白含量较高,能被斯氏狸殖吸虫病患者血清识别。2-DE电泳分离出的48个多肽斑点中,有10个含量较高,能被斯氏狸殖吸虫病患者血清识别,大部分为偏酸性的小分子多肽。     

Ultrastructural observation of spermatozoa and fertilization in Schistosoma japonicum

The ultrastructure of the sperm and the process of fertilization are described in Schistosoma japonicum. The sperm of S. japonicum has an elongated head and a single tail. The head measures 6.2 x 1.4 microm in average size. No acrosome is present. A mass of mitochondria locates in front of the nucleus. A layer of about 100-120 peripheral microtubules is parallel with the long axis of the head under plasma membrane. The nucleus is dense with some electron-lucent patches. The tail is a single flagellum with unique axoneme, which originates from a centriole. The structure of axoneme includes two types: 9 x 2 + in the main part of the flagellum, and 9 x 2 + 0 near the end of the flagellum. The sperm ultrastructure of S. japonicum is similar to that of other schistosomes, apart from the fact that two types of configuration coexisted in the same axoneme, and there is no striated root found in S. japonicum. The sperm differs distinctly from other Digenea. The aberrant ultrastructure of S. japonicum reflects that its evolution is far away from other genera in Digenea. Fertilization occurs at the posterior part of oviduct, in which region the oviduct wall lacks lamellae. Some cortical granules (CG) fuse with plasma membrane, and discharge their content on the surface of the fertilized ovum. The other CGs break down or degenerate in the cytoplasm. By the secondary mature division, the secondary oocyte finally divides to form a female pronucleus. During this period a male pronucleus also forms. The female and male pronucleus approach each other, come into contact in the central region and finally fuse to form a zygote. The function of CGs is discussed.     

Intracellular calcium increases with hyperactivation in intact, moving hamster sperm and oscillates with the flagellar beat cycle.

At some time before fertilization, mammalian sperm undergo a change in movement pattern, termed hyperactivation. There is evidence that hyperactivation offers an advantage to sperm for detaching from the oviductal mucosa, for penetrating viscoelastic substances in the oviduct, and for penetrating the zona pellucida. Hyperactivation is known to require extracellular calcium, but little else is known about the mechanisms by which calcium affects sperm movement. The calcium-sensitive fluorescent dye indo-1 was used to follow intracellular calcium levels ([Ca2+]i) in individual moving sperm. Sperm were loaded with 10 microM of the acetoxymethyl ester form of the dye and then rinsed. The dye was excited at 340 nm by using a filtered xenon stroboscope, and images at the 405-nm and 490-nm excitation maxima were simultaneously digitized at 30 per sec for 2.1 sec. [Ca2+]i was significantly higher in the acrosomal and postacrosomal regions of the head and in the flagellar midpiece (the principal piece could not be measured) in hyperactivated than in nonhyperactivated sperm (P < 0.0001). [Ca2+]i oscillations were detected in the proximal half of the midpiece that were identical in frequency to the flagellar-beat-cycle frequency in 12 of 17 hyperactivated sperm (median, 3.5 Hz). Rapid [Ca2+]i oscillations were also detected in the acrosomal and postacrosomal regions, as well as in the distal midpiece. Oscillations were not eliminated by dampening the flagellar bending with methyl cellulose. The [Ca2+]i oscillations detected in sperm are significantly more rapid than oscillations detected in other cell types.     

斯氏狸殖吸虫体壁神经细胞与神经纤维的观察

目的　探讨斯氏狸殖吸虫的体壁神经细胞和神经纤维的分布特点和相互间关系。方法　采用乙酰胆碱酯酶定位技术对斯氏狸殖吸虫的神经细胞和神经纤维进行显示。结果　斯氏狸殖吸虫前部体壁内神经纤维密集 ,以纵行神经纤维为主 ,虫体中部以相互交叉的斜行神经纤维为主。斯氏狸殖吸虫有两类神经细胞 ,即双极和多极神经细胞 ;虫体前部的纵行神经纤维上以多极神经细胞分布较多 ,中部较少 ,后部最少。双级神经细胞分布于神经节内、神经干内过神经干旁。在虫体的横截面上 ,神经细胞位于神经纤维下。结论　斯氏狸殖吸虫的前部体壁神经纤维密集 ,多极神经细胞较多 ,可能与虫体前部肌活动能力强有密切关系。     

华支睾吸虫亚显微结构的研究Ⅵ.精子的形成

华支睾吸虫精子的分化可以分为两部分,一是精细胞的发生,二是精子的形成,本文报道的为第二部分。精子的形成首先由精细胞核向细胞的游离端移动,然后出现细胞突起,在突起内可见中心粒和基体,此为分化区,继之胞质突起延伸形成中央突起,并在其两侧出现两个侧突起。细胞核变长,染色质浓缩,核纵断面呈丝状或版层状,核及线粒体分别移入分化区及中央突起内,随之中央突起与两侧突起融合,形成精子的杆状体,分化区的柱状体形成精子头部。最后精子自领状结构的水平与精细胞的胞质间桥分离,释放入睾丸腔内至此精子完全形成     

阴道毛滴虫超微结构的进一步观察

透射电镜观察虫体纵切面呈圆形或卵圆形。体前1/3有一椭圆形细胞核,在核与副基纤维的背侧有高尔基复合体。体前的毛基体复合体由鞭毛周管、管壁内“C”形盾结构及5个毛基体三部分组成。由毛基体分别发出4根前鞭毛及1根后鞭毛,同时还发出1根肋和2根副基纤维。轴柱分头、干、尾三部分,匙形轴头来自盾-轴连合处,围绕细胞核一侧向虫体后端延伸,至核末端扭转成筒状轴干,最后成针状伸出体外。     

Induction of an acrosomal process in Toxoplasma gondii: Visualization of actin filaments in a protozoan parasite

The invasive stages of Toxoplasma gondii, an Apicomplexan parasite, actively invade their host cells in an actin-dependent way. However, despite containing biochemically significant amounts of actin, actin filaments have never been observed in these parasites. Jasplakinolide, a membrane-permeable actin-polymerizing and filament-stabilizing drug, induced the polymerization of actin filaments at the anterior end of each tachyzoite in association with the conoid, where they formed, in many cases, a prominent membrane-enclosed apical projection reminiscent of acrosomal processes of invertebrate sperm. These jasplakinolide-induced filaments decorated with myosin subfragment 1, demonstrating unequivocally that they were indeed actin. Jasplakinolide-treated tachyzoites were unable to invade host cells, but once the drug was removed the parasites were able to enter host cells. Actin polymerization at the apical end of the parasite is consistent with the role of the apical end in host-cell invasion powered by a jackhammer-like extension and retraction of the conoid complex coupled to the secretion and rearward capping of surface proteins.     

Ultrastructural observation on the body wall of Pagumogonimus skrjabini

This paper deals with the ultrastructure of the body wall of adult Pagumogonimus skrjabini by transmission electron microscopy. Infected crabs were collected from Siyen, Hubei Province, and adult worms were obtained from the lungs of experimentally infected dogs 90 days post-infection. The normal structure of body wall of the P. skrjabini is composed of tegument, tegument cell, muscle muscle cell and protoplasmic tubules, all of which form together syncytium. The tegument contains external plasma membrane, tegument matrix and basal plasma membrane. The cell coat in fine granules is distributed over the whole external plasma membrane surface. The tegument matrix contains various secretory bodies, vesicles and mitochondria. The tegument cell is irregular in shape. Golgi complex, ribosome, autolysosome are seen in the cytoplasm. There are two layers of muscle, the external circular muscle and the inner longitudinal muscle layers. The nucleus of immature muscle cell has many heterochromatins, while the nucleus of mature muscle cell is large and round in shape. Mitochondria and glycogen granules are transmitted from muscle cell proper to the muscle by protoplasmic tubules (Figs. 1 approximately 7).     

Topographical Location of H—Y Antigen on Mouse Spermatozoa by Immunoelectronmicroscopy

H-Y (male) antigen was visually located on mouse sperm by electron microscopy, by use of the indirect hybrid antibody method and tobacco mosaic virus as the visual marker. Labeling was achieved by centrifugation of the sperm through a discontinuous gradient consisting of alternating layers of immune reagents and wash solutions. Treated sperm were examined topographically by preparation of platinum-carbon replicas. Antigen was located mainly on the acrosomal cap of the sperm head.