生物素-链霉亲和素免疫学法测定地高辛的研究

采用国产链霉亲和素直接包被塑料板孔,生物素标记抗体,建立的竞争酶联免疫吸附试验的方法测定血中地高辛浓度.其测定灵敏度为0.0964 μg/L,最低检测限为0.2251 μg/L,测定三份低、中、高浓度的血清标本,批内变异系数为8.9%,5.9%,2.4%;批间变异系数为15.8%,10.1%,9.2%,测定回收率在89.1%～107.22%之间,此法与FPIA方法相关良好(r=0.9488).     

过氧亚硝基-鲁米诺化学发光体系的改进

建立了一个测定过氧亚硝基阴离子（ONOO^－）化学发光的改进体系,测试了某些抗氧化剂清除ONOO^－的作用,其体系的组成和启动发光的程序如下：向pH 10.5碳酸缓冲液配的0.01 mol/L浓度NaN₃溶液通O₃ 30 s,取其800 μl原位注入含有100 μl水配样品和100 μl的0.001 mol/L鲁米诺溶液中,启动化学发光（chemiluminescence, CL）,立即测定每6秒的脉冲数（CP6S）,连续测定10～30次.根据实际需要,选其某一次的CL强度作为评判指标,对比抗氧化剂的活性.该发光体系灵敏、简便、且较稳定,最低可检测限为8.74 μmol/L的ONOO^－量,线性范围为8.74～74.04 μmol/L.批内变异系数3.35%（n=10）,批间变异系数5.52%（n=10）.测得维生素C(Vit.C)、茶多酚（EGCG）、原花菁素、硫脲皆有抑制CL,即清除ONOO^－的作用.     

以铁氰化钾为介体的苹果酸脱氢酶电极的研制

通过化学交联法将苹果酸脱氢酶 (MDH) 固定在玻碳电极表面(d=0.5cm),使用 N-甲基吩嗪甲基硫酸盐 (PMS) 和铁氰化钾为介体,间接地测定酶促反应中生成的还原辅酶 I(NADH).工作电位+350mV (vs.Ag/AgCl),L-苹果酸测定的线性范围为 25μmol/L—300μmol/L,响应时间小于60s,电极的使用寿命可达10d.并对电极的选择和重现性进行了讨论.     

比色法快速测定乳酸菌谷氨酸脱羧酶活力及其应用

基于Berthelot显色测定ω-氨基酸的反应 ,探讨了比色法快速测定谷氨酸脱羧酶活力的测定条件。结果表明 ,该方法灵敏度较高 ,重现性好 ,成本低 ,快捷 ,可替代氨基酸分析仪分析法。对乳酸菌谷氨酸脱羧酶提取液的适宜反应底物体系为 0.2mol LMacIlvaine ,pH4.7,内含 0.1mmol LPLP (5 磷酸吡哆醛 ) ,10mmol L底物L MSG (L 谷氨酸钠 )。 2 0 0 μL底物溶液和 1～ 1 0 0 μL酶液在 3 0℃反应 ,然后冰浴中加入 2 0 0 μ/L 0.     

纳米金生物条形码技术检测痕量二噁英类化合物

建立一种基于纳米金生物条形码技术的二噁英快速筛检方法.利用二噁英诱 导激活的芳香烃受体复合物,特异识别以纳米金为报告基团的二噁英反应探针,该 探针被释放后进行条形码放大,通过纳米金银染技术增强识别信号,并记录其吸光度值,从 而可以简单灵敏地快速筛检二噁英类化合物.在一定的反应时间和浓度范围内（10^-14～10^-10mol/L）,溶液的吸光度值与2,3,7,8 四氯二苯并二噁英（2,3,7,8-tetrachlorodibenzo-p-dioxin,TCDD）浓度之间呈正相关 ,方法检测限为0.01 pmol/L,变异系数为5%～8%.用纳米金生物条形码（Nano Barcod e,nanoparticle based bio barcode）方法和现有的生物分析方法（CALUX,chemical activated luciferase gene expression）分别测定TCDD标准品,并绘制剂量效应曲线.结 果表明,本方法灵敏度高,线性范围宽,重复性好.本研究纳米金生物条形码方法的检测灵 敏度高于CALUX将近5倍(EC₅₀分别为 4×10^-12 mol/L 和 2×10^{- 11} mol /L),检测限较CALUX降低了10倍（检测限分别为1×10^-14 mol/L 和1×10 ^-13 mol/L）,变异系数分别为5%～8%和15%～30%.     

林肯链霉菌丙氨酸脱氢酶的纯化和性质

采用硫酸铵分级沉淀、DEAE-纤维素52柱层析、亲和蓝柱层析和琼脂糖凝胶Sepharose6B柱层析的方法,分离纯化了林肯链霉菌丙氨酸脱氢酶,用聚丙烯酰胺凝胶电泳鉴定为单一组分。以凝胶过滤和聚丙烯酰胺梯度凝胶电泳测得该酶的分子量为170000,SDS-聚丙烯酰胺凝胶电泳测得其亚基分子量为42500,表明林肯链霉菌丙氨酸脱氢酶由四个相同的亚基组成。该酶加氨反应最适pH为9．0,脱氨反应最适pH为9．5,加氨反应和脱氨反应的最适温度均为50℃。加氨反应丙氨酸脱氢酶的表现米氏常数km值为：丙酮酸2．08×10－4mol／L,NH4+2．00×10-2mol／L,NADH2．38×10-5mol／L;脱氨反应的Km为：L-Ala1．43×10-2mol／L;NAD+6．67×10-5mol／L。     

高效毛细管电泳法直接测定红豆杉悬浮培养细胞中游离芳香族氨基酸

建立了红豆杉悬浮培养细胞中游离芳香族氨基酸的高效毛细管电泳的分离直接紫外吸收测定方法。在优化的实验条件下 ,以间苯二酚为内标 ,未涂层融硅毛细管 (75 μmi.d .,总长 37cm ,有效分离长度 30cm)为分离通道 ,40mmol/L磷酸缓冲液 (pH10 .46 )为电泳介质 ,3sec(0 .5 psi)压力进样 ,15kV恒压电泳 ,2 0 0nm检测 ,在 9min内三种氨基酸得到分离测定。色氨酸、苯丙氨酸和酪氨酸的线性范围分别为 2 .5 0～ 2 5 0 μmol/L(r =0 .996 2 ,RSD =2 .5 2 % )、1.2 5～ 2 5 0 μmol/L(r =0 .996 6 ,RSD =2 .12 % )和 2 .5 0～ 2 5 0 μmol/L(r=0 .9940 ,RSD =2 .93% ) ,苯丙氨酸回收率为 96 .8%± 1.37% ,酪氨酸回收率为 99.2 %± 3.0 4%。     

κ-银环蛇毒素敏感的烟碱受体激活引起的去甲肾上腺素释放参与烟碱诱导的长时程增强样反应

烟碱可以增强学习记忆功能,但其相关机制仍不清楚.海马长时程增强被认为是学习记忆的细胞机制.本研究室以往研究表明,当单脉冲的强度为诱发80%最大群体锋电位时,烟碱(10μmol/L)可以在海马CAI区诱导长时程增强样反应.本文通过细胞外记录离体海马脑片CA1区锥体细胞层群体锋电位,探讨烟碱诱导长时程增强样反应所涉及的烟碱受体亚型与相应的神经递质释放.结果显示,烟碱诱导的长时程增强样反应可以被美加明(mecamylamine,1 μmol/L)或κ-银环蛇毒素(κ-bungarotoxin,0.1μmol/L)阻断,但不被dihydro-β-erythtroidine(DHβE,10μmol/L)阻断.烟碱诱导的长时程增强样反应可以被普萘洛尔(propranolol,10μmol/L)阻断,但不被酚妥拉明(phentolamine,10μmol/L)或阿托品(atropine,10 μmol/L)阻断.以上结果提示,κ-银环蛇毒素敏感的烟碱受体激活引起的去甲肾上腺素释放参与烟碱诱导的海马CA1区长时程增强样反应.     

青杨脊虎天牛对植物源挥发物的EAG和行为反应

测定了青杨脊虎天牛Xylotrechus rusticus （L.）雌、雄成虫对其寄主杨树中的水杨醛（0.95 μmol/μL）和非寄主植物中0.3 μmol/μL的叶绿醇、0.4 μmol/μL的水芹烯和0.6 μmol/μL的 R 型α-蒎烯、S 型α-蒎烯、S型β-蒎烯、3-蒈烯、罗勒烯、香草烯和松节油等10种植物挥发性气味物质的触角电位（EAG）反应。结果表明,与对照相比,这10种植物挥发物多能引起成虫明显的EAG反应( P<0.05,P<0.01 ),其中雌虫对松节油、水杨醛、R 型α-蒎烯和 S 型α-蒎烯的EAG反应较强; 雄虫对 R 型α-蒎烯的EAG反应最强,松节油次之。根据雌虫对这10种挥发物EAG反应的强弱,进一步测定了雌虫对0.00006、0.0006、0.006、0.06、0.6、0.12 μmol/μL的松节油、R 型α-蒎烯、S 型α-蒎烯以及0.000095、0.00095、0.0095、0.095、0.95、0.19 μmol/μL的水杨醛的EAG和行为反应。结果表明,雌虫对松节油、水杨醛和 R 型α-蒎烯的EAG反应随气味物质浓度的增加而增加,水杨醛浓度增加到0.95 μmol/μL、松节油和 R 型α-蒎烯浓度增加到0.6 μmol/μL以后,EAG反应值趋于平稳;对 S 型α-蒎烯的反应随浓度的增加而呈线性增加。水杨醛浓度低于0.095时,对雌虫没有明显的定向作用( P>0.05 ),高于此浓度时表现为驱避作用( P<0.05 ); 松节油在浓度低于或等于0.6 μmol/μL时对雌虫表现为驱避作用,浓度为0.6时驱避效果最佳（ P<0.01 ）。雌虫对 R 型α-蒎烯和 S 型α-蒎烯没有明显的定向行为反应。     

黄鳝ISSR-PCR反应体系的建立及条件优化

以黄鳝基因组DNA为模板,采用正交试验设计方法,对各反应因子、引物退火温度和循环参数进行优化.建立了黄鳝的最适ISSR-PCR反应体系,25 μl反应体系中含2.5 mmol/ L Mg2+,250 μmol/ L dNTPs,0.25 μmol/ L 引物,1.0 U Taq DNA聚合酶,30 ng DNA模板.最佳反应程序:94℃预变性5 min;94℃变性40 s,48～57℃复性40 s(随引物而确定),72℃延伸1.5 min,循环次数40;72℃延伸10 min.利用所建立的ISSR反应体系,获得了清晰、重复性好、多态性高的DNA谱带.     

Recent advances in the study of Kaposi's sarcoma-associated herpesvirus replication and pathogenesis

It has now been over twenty years since a novel herpesviral genome was identified in Kaposi's sarcoma biopsies. Since then, the cumulative research effort by molecular biologists, virologists, clinicians, and epidemiologists alike has led to the extensive characterization of this tumor virus, Kaposi's sarcoma-associated herpesvirus(KSHV; also known as human herpesvirus 8(HHV-8)), and its associated diseases. Here we review the current knowledge of KSHV biology and pathogenesis, with a particular emphasis on new and exciting advances in the field of epigenetics. We also discuss the development and practicality of various cell culture and animal model systems to study KSHV replication and pathogenesis.     

The structure, reproduction and taxonomy of Vidalia and Osmundaria (Rhodophyta, Rhodomelaceae)

Comprises species occurring mostly in subtidal habitats in tropical, subtropical and warm-temperate areas of the world. An analysis of the type species, V. spiralis (Sonder) Lamouroux ex J. Agardh, a species from Australia, establishes basic characters for distinguishing species in the genus. These characters are (1) branching patterns of thalli, (2) flat blades that may be spiralled on their axis, (3) width of the blade, (4) primary or secondary derivation of sterile and fertile branchlets and (5) position of sterile and fertile branchlets on the thalli. Application of the latter two characters provides an important basic method for separation of species into three major groups. Osmundaria , a genus known only in southern Australia, was studied in relation to Vidalia , and its separation from the Vidalia assemblage is not accepted. Species of Vidalia therefore are transferred to the older genus name, Osmundaria. Two new species, Osmundaria papenfussii and Osmundaria oliveae are described from Natal. Confusion in the usage of the epithet, Vidalia fimbriala Brown ex Turner has been clarified, and Vidalia gregaria Falkenberg, described as an epiphyte on Osmundaria pro/ifera Lamouroux, is revealed to be young branches of the host, Osmundaria prolifera.     

New or rare chromosome counts in Artemisia L. (Asteraceae, Anthemideae) and related genera from Kazakhstan

Fifteen chromosome counts of six Artemisia taxa and one species of each of the genera Brachanthemum, Hippolytia, Kaschgaria, Lepidolopsis and Turaniphytum are reported from Kazakhstan. Three of them are new reports, two are not consistent with previous counts and the remainder are confirmations of very scarce (one to four) earlier records. All the populations studied have the same basic chromosome number, x = 9, with ploidy levels ranging from 2x to 6x. Some correlations between ploidy level, morphological characters and distribution are noted.     

肝癌中HBV和HCV基因和抗原的分布及意义

采用原位分子杂交方法检测HCV RNA及HBV X基因;采用免疫组织化学方法研究HCV核心抗原,非结构区C33c抗原及HBxAg在肝细胞肝癌中的定位及分布.结果表明(1)HCV RNA、HBV X基因在肝细胞肝癌组织检出率分别为40%(55/136)和82%(112/136).HCV RNA定位于癌细胞的胞浆内,阳性细胞呈散在、灶状及弥漫分布三种形式;HBV X基因在肝癌细胞中的分布呈胞浆型、核型及核浆型,阳性细胞也呈上述三种分布形式;(2)HCV C33c抗原、核心抗原在肝细胞肝癌中的阳性率为81%(133/164)及86%(141/164).C33c抗原定位于癌细胞及肝细胞的胞浆内;核心抗原既定位于癌细胞核中,又可定位于胞浆中.C33c抗原阳性细胞以灶状分布为主;而核心抗原阳性细     

Selection with two alleles and complete dominance

For a plant selection model with frequency-independent viabilities, fertilities and selfing rates, it is shown that apart from global fixation, for certain parameter combinations a protected polymorphism and facultative fixation (either allele may become fixed according to initial frequencies) may both occur. Facultative fixation requires different selling rates for the dominant and recessive type. Protection of the polymorphism requires resource allocation for male and female function. In this connection the problem of purely genetically caused population extinction is discussed.
For general frequency dependence and regular segregation, the chances for establishment of a completely recessive gene are compared to those of a completely dominant gene. It is proven that the process of establishment of the recessive gene, despite a fitness advantage, may be considerably endangered by drift effects if random mating prevails. The recessive gene may reach the same effectivity in establishment as a dominant gene, only if the recessive homozygote mates exclusively with its own type during the period of establishment.