冰冻切片法制作家兔眼球切片

用Ｓｚｅｎｔ－Ｇｙｏｒｇｙｒｉ液固定,明胶包埋,冰冻切片法制作 家兔眼球切片,获得了完整球壁的理想切片,其中睫状体,睫状突和睫状小带的形态结构更为理想。     

3种桉树组培苗不定根发生发育过程的解剖学观察

以巨尾桉‘GL9’、尾巨桉‘DH32-29’和尾边桉‘XF35’3种桉树无性系组培生根苗为材料,采用常规石蜡切片技术,对石蜡切片制作过程中的固定环节进行了优化,观察了桉树不定根的发育过程。结果表明:(1)采用FAA固定液固定材料,可获得染色清晰,组织完整的桉树根系切片。(2)‘GL9’和‘DH32-29’在生根诱导8d后生出不定根,生根类型为皮部生根;‘XF35’在生根诱导12d后生出不定根,生根类型为愈伤组织生根;‘GL9’不定根的根尖和根基处均有细胞旺盛分裂,‘DH32-29’不定根只在根尖有细胞旺盛分裂,‘XF35’不定根则只在其根基处有大量旺盛分裂的细胞。     

鬼笔属3种真菌的个体发育及形态观察

为揭示鬼笔属真菌子实体各器官的发育过程和形态特点,采用石蜡切片法对驯化栽培出菇的超短裙竹荪Phallus ultraduplicatus、冬荪P. dongsun和红托竹荪P. rubrovolvatus不同发育阶段的菇蕾进行切片显微观察,结合菌丝、孢子和子实体形态比较3个种的个体发育及形态特点。结果表明：3个种的菌丝和孢子形态差异不明显,超短裙竹荪子实体的菌裙质地较薄、长约4 cm、带有不规则形状的网孔;红托竹荪的菌裙质地较厚、长约7 cm、带有圆形网孔;冬荪仅有1层薄菌幕。3个种的菇蕾内部发育顺序一致,最先发育的都是胶质腔,接着是子实层、菌柄腔、菌盖、菌柄、菌裙（菌幕）;发育过程中菌丝形态经历从疏松到紧密、从交织状到组织状的变化过程;各器官发育完成的时间不同,在菇蕾直径相同的条件下,冬荪发育最快,在菇蕾直径为2 cm时,各器官基本发育完成,红托竹荪在菇蕾直径为3 cm时发育完成,超短裙竹荪的菇蕾发育最慢,在直径达到4 cm时各器官才能发育完成。为鬼笔类真菌的分类研究和亲缘关系探索提供了依据。     

木耳属真菌rDNA特异性扩增片段的RFLP研究

对木耳属8个种25个菌株的ITS和28SrDNA5’端两个区域分别进行了PCR扩增和限制酶切片段长度多态性（RFLP）研究。ITS－RFMP研究结果表明,HaeⅢ可将黑木耳与其它种区分开,MspⅠ可将盾形木耳、角质木耳、琥珀木耳和黑木耳4个种区分开,而供试的HaeⅢ、TaqⅠ、HinfⅠ和MaPⅠ这四种限制酶均不能将皱木耳、大木耳、网脉木耳及毛木耳4个种区分开,表明它们之间的亲缘关系较近;结果还表明,ITS—rDNA拷贝在毛木耳和琥珀木耳种内是异质性的,而在黑木耳种内是同质性的。285rDNA-RFLP研究结果表明,供试的4种限制酶中,仅MspⅠ可将盾形木耳和角质木耳区分开,而不能将其它种区分开,这显示了28SrDNA序列在木耳属不同种间的保守性。     

五指山猪生物学特性、易地繁育及遗传多样性研究

五指山猪是我国濒临灭绝的珍稀品种,为保护和利用这一特有种猪基因资源,我们开展了易地繁育和生物学特性及遗传多样性的研究。通过对其生长发育规律、繁殖生理、血液生理生化、遗传标记、DNA指纹图谱等项的观察和测试,发现五指山猪具有体型小、性成熟早、耐近交繁殖、遗传性较稳定、肉质好等特点。白细胞分类中淋巴细胞占77％,白细胞抗原(SLA) 血清学分型中,与其它品种之间存在有差异性。 用人源小卫星探针33.6和鸡小卫星探针cMS18,对五指山猪及长白猪和枫泾猪进行DNA指纹图比较分析,发现五指山猪的DNA指纹图相似系数,大大高于其它已研究过的正常体型猪种,说明它经历过较高程度的近交;以猪的生长激素（GH）cDNA为探针,用EcoRI、EcoRV、PstI等酶切基因组DNA进行RFLP分析时,五指山猪均出现了一条深色带,而长白猪和枫泾猪未发现。表明五指山猪在生长激素位点上与正常体型猪种亦存在差异。     

八角莲组织培养的器官发生途径研究

为探究小檗科植物八角莲组织培养的器官发生方式,该研究以八角莲离体叶片、叶柄在MS培养基上诱导产生的愈伤组织、不定芽、不定根为对象,用连续石蜡切片技术分析八角莲组织培养的器官发生途径。结果表明:八角莲愈伤组织形成的解剖学特征是靠近表皮的薄壁细胞经激素刺激恢复分裂能力,继续培养形成拟分生组织。拟分生组织可形成许多分化中心。通过对八角莲组织培养产生的不定芽细胞组织学观察发现芽原基起源于愈伤组织外侧的几层薄壁细胞,芽原基背离愈伤组织中央生长形成不定芽,故八角莲脱分化形成的芽起源方式为外起源。而八角莲的根原基起源于组织深处髓部薄壁细胞和部分维管形成层细胞,进而形成类似球形或楔形并朝韧皮部突起的根原基轮廓,根原基继续发育会突破表皮生成不定根,起源方式为内起源。八角莲离体再生途径为器官发生型,在组培苗生长过程中先诱导形成不定芽,再诱导形成不定根,在愈伤组织上形成维管组织将不定芽和不定根连接成完整植株。     

中国人胃癌染色体17q21区域基因遗传不稳定性研究

To investigate microsatellite instability (MSI) and loss of heterozygosity (LOH) of locus D17S396, D17S579 and D17S855, and their effect on the expression of nm23H1 and BRCA1 of gastric cancer, which would provide experimental basis for clinical treatment and prognosis analysis of gastric cancer. DNA was extracted from paraffin-embedded materials. Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) was used to analyze MSI and LOH. Expression of nm23H, and BRCA1 was detected by Envision immuno-histochemistry and Leica-Qwin computer imaging techniques. In the forty cases of gastric cancer, the frequency of MSI, LOH and nm23H1 protein were 20.00%, 17.50% and 55.00% respectively at locus D17S396, while at locus D17S579, the frequency of MSI, LOH and BRCA1 protein were 22.50%, 15.00% and 37.50% respectively; at locus D17S855, the frequency of MSI, LOH and BRCA1 of thirty-seven cases were 18.92%, 18.92%, 37.84% respectively. In tumor node metastasis (TNM) staging, at locus D17S396, D17S579 and D17S855, MSI in stages I + II appeared more frequently than that in stages III + IV, while LOH appeared the contrary tendency. In the group of metastasis of gastric cancer, MSI had a less frequency (5.00%) than that with no metastasis (35.00%, P < 0.05) at locus D17S396, but LOH appeared more frequently (30.00%) than that with no metastasis (5.00%, P < 0.05). At locus D17S579, MSI had an increasing tendency with the degree of tumor differentiation (50.00% in high differentiation cases, 20.00% in middle differentiation cases, and 0% in low differentiation cases, P < 0.05). The frequency of nm23H1 and BRCA1 protein in stages TNM I + II was higher than that in stages TNM III + IV; and that in higher differentiation cases was higher than in poor differentiation cases. The frequency of nm23H1 protein in the group of metastasis (30.00%) was less than that with no metastasis significantly (80.00%, P<0.01). The frequency of nm23H1 protein in the group positive to MSI (87.50%) was higher than that in the group negative to MSI (46.88%, P < 0.05). However, nm23H1 protein in group positive to LOH (14.29%) was lower than that in the group negative to LOH (63.64%, P < 0.05). The frequency of BRCA1 protein in the group positive to MSI (66.67%) was more than that in the group negative to MSI (29.03%, P < 0.05). The results of experiments indicate that MSI and LOH may separately control the development of sporadic colon cancer with different pathways. MSI may be an early period molecule marker for sporadic colon cancer, enhanced expression of nm23H1 protein can effectively inhibit colon cancer metastasis and improve prognosis of sporadic colon cancer patients. By comparison, LOH mostly arises in the late period of sporadic colon cancer and endows a high aggressive and poor prognostic phenotype. nm23H1 protein could effectively restrain gastric cancer metastasis and development; and BRCA1 protein could restain tumor from becoming lower differentiation.     

异色瓢虫视觉系统中5-HT阳性神经元的分布

本文运用树脂石蜡（Colophony-Paraffin,CP;专利号：ZL98125709．7）组织包埋切片技术,结合免疫组织化学链酶菌抗生物素蛋白一过氧化物酶（Streptavidin-Peroxidase,SP）双染法,对异色瓢虫视觉系统中5-羟色胺（5-HT）能神经元的分布进行了初步研究。结果显示,异色瓢虫视觉系统的结构及5-HT免疫反应系统相对比较特殊。5-HT阳性神经元胞体数目较少,染色显著,并聚集成群。根据胞体定位、细胞形态及轴突走向,可大体分为5群,其中包括1群呈弱反应的光感细胞。5-HT阳性膨大纤维支配所有的视神经纤维网,并呈柱状或分层排列模式。结果表明5-HT作为经典的神经递质在昆虫的视觉信息处理过程中可能发挥重要的调节作用,且主要以远距离的广域神经调节模式为主,并在特定区域和GABA有伴随现象。此外,昆虫视觉系统中5-HT的含量还可能与其明暗适应的生理调节方式具有相关性[动物学报51（5）：912—918,2005]。     

昆虫嗅觉神经的计算机三维重建

基于激光扫描共聚焦显微镜平台的计算机三维重建在昆虫嗅觉神经研究中发挥了重要作用。对经荧光标记的神经组织采集系列光学切片并进行三维重建,在双翅目、鳞翅目、膜翅目、蜚蠊目昆虫中均有进展。触角叶是昆虫的初级嗅觉中心,触角叶的解剖学图谱是识别不同种和雌雄虫间嗅球体特定功能的先决条件。了解构成嗅觉传输途径的主要神经元的形态和空间关系是理解气味信息在中枢神经系统编码的基础。三维重建昆虫的嗅觉神经,对于探讨昆虫嗅觉在其寄主选择、觅食以及寻找配偶等行为中的作用具有非常重要的意义。     

介绍一种简单适用的胸腹水脱落细胞石蜡包埋法

常规胸腹水脱落细胞检测多采用直接涂片法,该法简便、快速,在临床上得以广泛应用。但在实际工作中,我们发现直接涂片常出现细胞重叠,堆积,厚薄不均匀,细胞结构及背景不清晰,细胞量少,细胞脱落等现象,往往导致癌细胞检出率低。特别是某些需要通过免疫细胞化学进行鉴别诊断的病例,如转移性腺癌细胞与问皮源性细胞,诊断前一般并未预留涂片,在进行HE染色涂片观察后,