几种濒危植物及其近缘类群总DNA的提取与鉴定

用低ｐＨ 介质,高盐沉淀蛋白质方法成功地从银杉（Ｃａｔｈａｙａ ａｒｇｙｒｏｐｈｙｌｌａ Ｃｈｕｎ ｅｔＫｕａｎｇ）、矮牡丹（Ｐａｅｏｎｉａ ｓｕｆｆｒｕｔｉｃｏｓａ ｖａｒ． ｓｐｏｎｔａｎｅａ）、南川升麻（Ｃｉｍ ｉｃｉｆｕｇａ ｎａｎｃｈｕａｎｅｎｓｉｓＨｓｉａｏ）、裂叶沙参（Ａｄｅｎｏｐｈｏｒａ ｌｏｂｏｐｈｙｌｌａ）的同属种泡沙参（Ａ． ｐｏｔａｎｉｎｉｉ）等植物中提取和部分纯化了细胞总ＤＮＡ,并对其产率、质量和纯度作了鉴定。此方法的关键是用了一个低ｐＨ提取介质,它能有效防止组织破碎及沉淀大量材料时的电离化作用及酚化合物的进一步氧化。所得ＤＮＡ 不需经氯化铯梯度离心或柱层析,直接可用于限制性片断长度多态性（ＲＦＬＰ）及随机扩增的ＤＮＡ多态性（ＲＡＰＤ）等分子水平的遗传标记。为检测濒危植物的遗传多样性提供了一套迅速、简便和可靠的技术方案     

Systemic amyloidosis in transgenic mice carrying the human mutant transthyretin (Met 30) gene

To analyze the pathologic processes of amyloid deposition in type I familial amyloidotic polyneuropathy (FAP), mice were made transgenic by introducing the human mutant transthyretin (TTR) gene(MT-hMet 30). An inbred strain of mouse, C57 BL/6, was chosen. Transgenic mice were killed using ether anesthesia at 3-mo intervals up to 24 mo after birth. In these transgenic mice, amyloid deposition started in the gastrointestinal tract, cardiovascular system, and kidneys and extended to various other organs and tissues with advancing age. The pattern of amyloid deposition was similar to that observed in human autopsy cases of FAP, except for its absence in the choroid plexus and in the peripheral and autonomic nervous systems. We extracted the amyloid fibrils from kidneys of these mice with a human mutant TTR gene and analyzed them immunochemically and electronmicroscopically. Deposited amyloid was shown to be composed of human mutant TTR and mouse serum amyloid P component. Amyloid fibril from transgenic mice was morphologically and immunohistochemically similar to that of human FAP. The most striking pathologic feature of the transgenic mice was the absence of amyloid deposition in the peripheral and autonomic nervous tissues. Thus, other intrinsic factors may be involved in amyloid deposition in the nervous tissues of human FAP.     

大血藤科植物的分类学研究

作者对大血藤科植物的花性、叶片内部结构形态、花粉形态、染色体核型及过氧化物酶和脂酶同工酶进行了比较研究,结果表明：１）大血藤科植物外形上的两性花,其雄蕊的形态退化、花药始终不开裂,为功能上的雌花,因此其花为单性,同株,同序或异序;１２）首次指出大血藤属植物的染色体数为２ｎ＝２ｘ＝２２,属小型染色体,与木通科植物２ｎ＝２ｘ＝３２、３０、２８的染色体数明显不同,支持Ｓｔａｐｆ（１９２６）将其从木通科分出另立为科的观点;３）单叶和复叶可出现于同一植株上,而且在叶片内部结构、花粉形态、染色体核型、过氧化物酶及脂酶同工酶等性状上,大血藤与单叶血滕间均无实质性的差异,因此将单叶血藤归并于大血藤中。     

鸡马立克氏病病毒B抗原片段在大肠杆菌中表达

鸡马立克氏病病毒ＢａｍＨＩ基因文库Ｉ３质粒中含有编码Ｂ抗原膜外Ｄｏｍａｉｎ的ＤＮＡ序列。经ＳｃａＩ和ＳｐｈＩ双酶酶解Ｉ３质粒,分离获得７６４ｂｐＤＮＡ片段,并克隆进Ｍ１３ｍｐ１９中。ＤＮＡ序列测定分析表明克隆片段为ＭＤＶ－Ｂ抗原基因的４９４－１２５８ｂｐ部分序列。进一步分离ＮｃｏＩ－ＨｉｎｄＩＩＩ部分基因片（５３０ｂｐ）,克隆于ＰＬＰｒｏｍｏｔｅｒ控制下的含有修饰型ｃＩｔｓ８５７基因的表达载体中,     

Regulation of vasoactive intestinal peptide expression in sympathetic neurons in culture and after axotomy: The role of cholinergic differentiation factor/leukemia inhibitory factor

Vasoactive intestinal peptide (VIP) expression increases in sympathetic neurons when they are grown in dissociated cell or explant cultures and when they are axotomized in vivo. In dissociated cell culture, the magnitude of the VIP increase was reduced when nonneuronal cells were removed and medium conditioned by ganglionic nonneuronal cells increased VIP in neuron-enriched cultures. Antiserum Against cholinergic differentiation factor (also leukemia inhibitory factor; CDF/LIF), but not against ciliary neurotrophic factor, immunoprecipitated this activity. Medium conditioned by sympathetic ganglion explants also contained a VIP-stimulatory molecule that was immunoprecipitated by CDF/LIF antiserum, and CDF/LIF antiserum partially blocked VIP induction in explants. CDF/LIF mRNA was increased in dissociated cell cultures, in ganglion explants and in vivo after axotomy. Our results suggest that CDF/LIF released from ganglionic nonneuronal cells plays an important role in regulating VIP after axotomy. 1994 John Wiley & Sons, Inc.     

从一名国内感染的艾滋病人分离人免疫缺陷病毒（HIV）

从一名国内感染的艾滋病人采血,分离其外周血单核细胞（ＰＭＣｓ）。首先与正常的ＰＭＣｓ共培养,４周后检测其ＨＩＶ－１ｐ２４抗原（ＥＬＩＳＡ）达到峰值。用此时的细胞及其上清分别感染Ｊｕｒｋａｔ－ｔａｔ、ＣＥＭ、ＭＴ４细胞,可很快地在这三株细胞中检测到ＨＩＶ生长,ＨＩＶ在Ｊｕｒｋａｔ－ｔａｔ细胞中生长情况最好,同时用病人血清直接感染Ｊｕｒｋａｔ－ｔａｔ和ＭＴ４细胞,４周后检测其细胞上清ＨＩＶ－１ｐ２４抗原（ＥＬＩＳＡ法）为阳性,但ＯＤ值很低（约为ＰＭＣ共培养组的一半）。用病人的少量全血与正常ＰＭＣｓ共培养,得到的结果与分离病人ＰＭＣｓ法相近。应用间接免疫荧光法（ＩＦＡ）、免疫酶法（ＩＥＡ）、蛋白印迹法及ＨＩＶ－１ＰＯｌ基因和Ｅｎｖ基因特异引物的聚合酶链反应（ＰＣＲ）等证实为ＨＩＶ－１病毒。分离的ＨＩＶ在Ｊｕｒｋａｔ－ｔａｔ细胞中连续传代,细胞被感染后２－３天即出现以大量融合细胞为主的细胞病变,感染后７－１０天细胞几乎全部死亡。病毒在连续传代过程中的生长特征及致细胞病变特征不变。此病毒命名为ＣＡ－２毒株。     

高脂血症兔球结膜微循环的改变

本实验用胆固醇粉喂家兔,造成实验性高脂血症,活体观察球结膜微循环的形态、流态及血管周围状态等１５项指标,并用球结膜微循环综合定量评价方法计算了综合积分值。处死后取球结膜进行组织学检查。结果表明：高脂血症家兔球结膜微循环有明显改变,主要为微血管形态异常、血流减慢、红细胞聚集、白微栓、静脉壁上有白色斑块等改变。球结膜微循环综合积分值明显增加,高于实验前。球结膜组织学检查发现上皮层下有泡沫细胞、血管壁增厚、有空泡、静脉内有血栓。虹膜睫状体内除有多数泡沫细胞外,还有胆固醇的菱形结晶。上述改变表明,高脂血症兔球结膜微循环有明显改变。血液有高凝、高聚倾向。黑茶中的茶色素有抗凝、抑制血小板聚集、促进纤溶等作用,口服黑茶可改善高脂血症兔球结膜微循环障碍。     

气相色谱法测定梭菌酸性代谢产物

本文报告用气相色谱法对８株梭菌代谢产物中的挥发性及非挥发性有机酸进行测定。采用程序升温方法,样品中各种成份分离较好,所需样品量少,操作方便,样品分析结果与标准菌株及文献对照比较,为梭菌的分类鉴定提供了实验依据。     

流行性出血热免疫球蛋白的研制

本文首次报告采用纯化灭活流行性出血热（ＥＨＦ）Ⅰ型疫苗免疫健康献血员,采集ＥＨＦ抗体高滴度的血浆,用低温乙醇法及盐析法分离纯化三批ＥＨＦ免疫球蛋白。结果表明：（１）采用０,１,３,４（月）或０,１,４,５（月）免疫程序,疫苗剂量１～２ｍｌ（０．１５～０．３０ｍｇ蛋白）,受免献血员血清平均抗体滴度可达１∶４０６（ＥＬＩＳＡ）或１∶１１２（ＲＰＨＩ）。（２）通过生化检定,三批制品的电泳纯度为９７．０５％,９６．８４％,９９．２６％;ＩｇＧ单体和二聚体含量为８９．５５％,９１．３０％和９８．２１％。（３）用空斑抑制中和试验及免疫印染试验证明所纯化的免疫球蛋白具有抗ＥＨＦ病毒特异性。（４）三批ＥＨＦ免疫球蛋白的效价测定,结果为ＥＬＩＳＡ滴度≥１∶５１２,ＲＰＨＩ滴度≥１∶１０２４,ＰＲＮＴ（中和抗体）滴度１∶４０。按１６％蛋白计,ＥＨＦ免疫球蛋白的效价可达原料血浆的１０倍以上。（５）无菌、安全、毒性及热原质试验检定结果,全部通过《中国生物制品规程》要求。