Active site of trichosanthin acting as a ribosome-inactivating protein

天花粉蛋白核糖体灭活活性部位的定位方法：羟胺特异裂解天花粉蛋白唯一ＡｓｎＧｌｙ肽键．制备性凝胶电泳获ＨＡＴｆ１和ＨＡＴｆ２二片段．免疫印迹确定天花粉蛋白上不同表位并筛选抗体．兔网织红无细胞系统测定天花粉蛋白及片段对蛋白合成的抑制活性．结果：ＨＡＴｆ１和ＨＡＴｆ２纯度各达９６６％和８０５％．ＨＡＴｆ１保留完整天花粉蛋白的抑制活性．第１４号和第１６号抗天花粉蛋白单抗与二片段显示不同免疫反应性，并用于封闭试验．第１４号单抗能封闭天花粉蛋白及ＨＡＴｆ１活性，而第１６号单抗则否．结论：天花粉蛋白抑制蛋白质生物合成的活性部位位于ＨＡＴｆ１侧，近二部分交界．     

广东省内常见乙肝检测ELISA试剂的抽样调查

广东省临床检验中心组织广州市九间三级甲等医院的免疫检验专家，抽样调查了九个有生产批文的试剂厂家出品的乙肝检测ＥＬＩＳＡ试剂。结果显示：不同厂家试剂的灵敏度相差１－５个稀释度，精密性为１．２８～６６．３％；九厂家试剂的平均特异性为８９．４～１００％、敏感性为７５～９４％、符合率为８６．５～９５．９％；五种品名试剂的平均特异性ＨＢｓＡｇ９８．８％、抗－ＨＢｓ９７．０％、ＨＢｅＡｇ９９．１％、抗－ＨＢｅ９６．２％、抗－ＨＢｃ８６．５％、平均敏感性ＨＢｓＡｇ８５．５％、抗－ＨＢｓ７８．６％、ＨＢｅＡｇ９０％、抗－ＨＢｅ７０．２％、抗－ＨＢｃ９３．１％，平均符合率ＨＢｓＡｇ９０％、抗－ＨＢｓ８９．９％、ＨＢｅＡｇ９５．１％、抗－ＨＢｅ８６．１％、抗ＨＢｃ９０．６％。     

没食子酸酯对羟自由基损伤抗凝血酶Ⅲ的保护作用

ＣｕＣｌ＿２／抗坏ＭＫ体系产生的羟自由基（·ＯＨ）在离体条件下可抑制血浆及纯品抗凝血酶Ⅲ（ＡＴ－Ⅲ）活性，诱发ＡＴ－Ⅲ纯品解聚，这种损伤呈明显的量效。时效关系。且血浆稀释度越高。ＡＴ－Ⅲ损伤越重。０．１－０．６ｍｍｏｌ·Ｌ￣（－１）没食子酸乙酯。丙酯和异丁酯抑制血浆ＡＴ－Ⅲ氧化呈量效关系。０．６ｍｍｏｌ·Ｌ￣（－１）时，上述三种酯对ＡＴ－Ⅲ氧化的抑制率依次为４８．５％，３９．１％和４０．０％，比较结果表明。这些酯与另三种抗血栓药抑制ＡＴ－Ⅲ纯品解聚的作用强度为川芎嗪＜阿魏酸钠＜儿茶精＜没食子酸酯．     

高效液相色谱法测定盐酸氟桂利嗪血浆浓度及其药物动力学

以高效液相色谱法测定血浆中的盐酸氟桂利嗪浓度。色谱条件：紫外检测波长２１０ｎｍ；柱ＳｐｈｅｒｉｓｏｒｂＣ８１５０×４．６ｍｍ；流动相：甲醇（８０％）∶水（１５％）∶（０．０５ｍｏｌ·Ｌ－１ＮＨ４Ｈ２ＰＯ４＋０．０２５ｍｏｌ·Ｌ－１Ｈ３ＰＯ４）缓冲液（５％）。最低检测限１０ｎｇ·ｍｌ－１。药物动力学为一室模型，其参数（Ｔ１／２（Ｋ）２．３８～２．６４ｈ，Ｔｍａｘ２．３０～２．４３ｈ，Ｃｍａｘ１３５．１３～１４３．９６ｎｇ·ｍｌ－１。     

rhIL-6大肠杆菌高效表达的影响因素及其产品中试的研究

研究自构建的重组人白细胞介素－ ６（ｒｈＩＬ－６）基因工程菌株高效表达的影响因素及其ｒｈＩＬ－６产品中试工艺。方法：①将ｒｈＬ－６－ＰＢＶ重组质粒转化至ＲＲＩ、ＪＭ１０３和 ＤＨ５ ａ不同宿主菌中,在Ｍ９ＣＡ和 ＬＢ培养基中,４２℃诱导培养不同时间（３～６ ｈ）,观测 ｒｈＩＬ－６不同表达效率。②采用 １０ Ｌ发酵罐诱导培养,经破菌－洗包－溶包初步纯化后,再经三步柱层析纯化,提取ｒｈＩＬ－６。结果：①ｒｈＩＬ－６在ＤＨ５ ａ大肠杆菌中（ＳＤＨ－９４５株）表达效率高;可达 ３９％,用 Ｍ９ＣＡ培基,４２℃诱导培养 ５ｈ可使表达效率提高到４１％,②ＳＤＨ－９４５菌株在５批 １０Ｌ Ｍ９ＣＡ发酵诱导培养 ５ ｈ,ｒｈＩＬ－６表达效率可达 ３５． ８７±２． ６５％。经初步纯化后,ｒｈＩＬ－６的纯度达 ７４． ８７±３． ６８％,再经三步柱层析后,纯度可达 ９５％以上,比活性达 ４ × １０８ Ｕ／ｍｇ,总得率稍低可达 ２３．１％．结论：①ＳＤＨ－９４５工程菌株在 Ｍ９ＣＡ培养基巾,４２℃诱导培养 ５ ｈ,ｒｈＩＬ－６表达效率最佳。②１０ Ｌ发酵和纯化工艺可获高效价、高比活性、高纯度的ｒｈＩＬ－６合格生物制品。     

4,5—二氢—6—[（苯乙酰基—哌嗪基）苯基]—5—甲基—3（2H）—哒…

４，５－二氢－６－［（苯乙酰基－哌嗪基）苯基］－５－甲基－３（２Ｈ）－达嗪酮（ＳＭⅡ４）０．１－２．５μｍｏｌ．Ｌ＾－＾１能使血小板激活因子（ＰＡＦ）及血栓素Ａ２类似物Ｕ４６６１９诱导的兔血小板聚集剂量一效应曲线左移且最大反应降低。其ｐＤ２分别为６．０±ｓ０．４及６．１±ｓ０．３．ＳＭⅡ４还能抑制ＡＤＰ，花生四烯酸（ＡＡ）及Ｕ４６６１９诱导的人血小板聚集，其ＩＣ５０分别是１．２，１．３及１．６．．     

肿瘤转移抑制基因nm23mRNA在大肠良,恶性病变中的表达及意义

为探讨肿瘤抑制基因ｎｍ２３－Ｈ１ｍＲＮＡ表达对鉴别大肠良恶性病变的意义，应用原位分子杂交技术对９３例大肠癌、６２例癌旁粘膜、６例大肠腺瘤、７例增生性息肉、９全慢性结肠炎及５例正常在肠组织中ｎｍ２３－Ｈ１ｍＲＮＡ的表达进行了研究。结果明显，上述在大肠组织中ｎｍ２３－Ｈ１ｍＲＮＡ的阳性表达率分别为７９．６％、７５．８％，８３．３％、８５．７％、７７．７％和８０．０％，各组间差异无显性，而大肠癌中ｎｍ     

知母宁清除自由基作用的量子化学研究(英文)

研究知母宁的　酮结构是否有利于提高其清除自由基活性并探讨知母宁酚羟基活性的顺序．方法：用半经验量子化学方法ＡＭＩ计算出知母宁不同状态的△ＨＯＦ值（母体化合物与其自由基生成热之差）和电子自旋密度分布,以△ＨＯＦ值作为衡量知母宁清除自由基活性的理论指标,观察结构因素对△ＨＯＦ值的影响．结果：计算得出不同酚羟基的△ＨＯＦ值为 Ｏ５－Ｈ６： １３９．０９ ｋＪ·ｍｏｌ－１; Ｏ４ －Ｈ５： １４１．４６ ｋＪ· ｍｏｌ－１; Ｏ２－Ｈ２： １８５．１４ ｋＪ·ｍｏｌ－１; Ｏ１－Ｈ１： １９５．７１ ｋＪ·ｍｏｌ－１,同时得到自由基的自旋密度分布．结论：知母宁的　酮结构使Ｃ环对Ｂ环有较强的钝化作用,降低了Ｂ环酚羟基清除自由基的活性．知母宁酚羟基活性的顺序为Ｏ５－Ｈ６＞Ｏ４－Ｈ５＞Ｏ２－Ｈ２＞Ｏｌ－Ｈｌ。     

洛之达治疗高脂血症的临床观察

高血脂症３４例，服洛之达２０ｍｇ，ｑｎ，疗程８周，服药８周后与服药前比较，血清ＬＤＬ－Ｃ，（ＴＣ－ＨＤＬ－Ｃ）／ＨＤＬ－Ｃ及ＡｐｏＢ分别下降２６．８％，２４．１％及２２．２％，ＴＣ下降１９．５％，ＴＧ下降１４．５％。血清ＨＤＬ－Ｃ，ＡｐｏＩＡＩ及ＡｐｏｉＡＩ／ＡｐｏＢ分别上升６．７％，１０．１％及４０．０％。     

HOAP方案治疗急性非淋巴细胞白血病疗效分析

用ＨＯＡＰ方案治疗急性非淋巴细胞白血病（ＡＮＬＬ）６６例，完全缓解（ＣＲ）４７例（７１．２％），部分缓解（ＰＲ）６例（９．１％），总缓解率８０．３％ 。对４７例ＣＲ患者随访观察，存活１年以内的９例（１９％），１￣２年的９例（４７％），２￣３年的３例（６．４％），３￣６年的９例（１９．１％），６￣１３年的４例（８．５％）。通过随访分析显示诱导缓解有效，坚持定期复查系统维持治疗者预后较好，存活期长。     

C16 and C17 substituted derivatives of pregnenolone and progesterone as inhibitors of 17alpha-hydroxylase-C17, 20-lyase: synthesis and biological evaluation

17 alpha-hydroxylase-C17, 20-lyase (P450 17, CYP 17) is a key enzyme in androgen biosynthesis and a target for the treatment of prostate cancer. In order to find novel inhibitors for this enzyme, several compounds bearing different moieties able to complex with the heme iron located in the active site of the enzyme were synthesized. The moieties were introduced into the 16-position of pregnenolone and progesterone. Their inhibitory activities toward human and rat CYP 17 were determined and compared to the activities of the corresponding 17-substituted compounds. It became apparent that the 16-substituted compounds were less active than the parent compounds: they were either moderate or poor inhibitors of the target enzyme. Tested for inhibition of human 5 alpha-reductase 1 and 2--a target for the treatment of benign prostatic hyperplasia (BPH)--the title compounds showed some inhibitory activity.     

Design and screening of antisense oligodeoxynucleotides against PAI-1 mRNA in endothelial cells in vitro

AIM: To design and screen antisense oligodeoxynucleotides (ASODNs), which inhibit type-1 plasminogen activator inhibitor (PAI-1) expression in human umbilical vein endothelial cells (HUVEC) in vitro. METHODS: Twenty seven ASODNs against different sites of PAI-1 mRNA were designed and transfected to HUVEC by lipofectin in vitro. The effects of ASODNs on PAI-1 antigen, PAI-1 activity and PAI-1 mRNA expression were detected by ELISA, amidolytical assay and RT-PCR, respectively. RESULTS: Transforming growth factor beta1 (TGF-beta1)-treated HUVEC increased the expression of PAI-1 compared with the normal HUVEC. Five among twenty seven designed ASODNs were effective in inhibiting the increase in PAI-1 antigen and PAI-1 activity in a dose-dependent manner after 48-h transfection. In particular, ASODN 14 (AO14) exhibited the best inhibitory effect. The control sequences of AO14, including sense, scramble, and mismatch sequences, did not significantly inhibit PAI-1 activity. It was revealed that the inhibitory efficacy of AO14 was in a sequence-specific manner. RT-PCR showed that ASODN 1, 7, 8, 14, and 15 decreased PAI-1 mRNA expression induced by TGF-beta1 and AO14 showed the best inhibitory effect. CONCLUSION: ASODN 1, 7, 8, 14, and 15, among twenty seven designed ASODNs against PAI-1 mRNA, significantly decreased PAI-1 antigen and PAI-1 activity induced by TGF-beta1 in a dose-dependent manner in HUVEC in vitro. AO14 showed the best inhibitory effect on PAI-1 expression in a sequence-specific manner. The results of RT-PCR indicated that inhibitory effects of ASODNs on PAI-1 biosynthesis occurred at the mRNA level. Four among five effective target sites of ASODNs located at the translation initiation site or within the translation area of PAI-1 mRNA, suggesting that these sites may be promising sites for the design of effective ASODNs.     

Medicinal plants of surinam.

From ASPIDOSPERMA EXCELSUM, rootbark, which has previously been shown to exhibit antimicrobial activity, 6 antimicrobial active alkaloids were isolated and identified: 11-methoxytubotaiwine, ochrolifuanine A, tetrahydrosecamine, 16-demethoxycarbonyltetrahydrosecamine, 16-hydroxytetrahydrosecamine and 16-hydroxy, 16-demethoxycarbonyltetrahydrosecamine. The minimum inhibitory concentration was determined for some of the alkaloids, using BACILLUS SUBTILIS as test organism. Didemethoxycarbonyltetrahydrosecamine was also shown to exhibit antimicrobial activity. No activity against gram negative bacteria could be found for the alkaloids by means of biograms. The site of demethoxycarbonylation in demethoxycarbonyltetrahydrosecamine was established as C-16. The structures of the two new secamine type alkaloids, 16-hydroxytetrahydrosecamine and 16-hydroxy,16-demethoxycarbonyltetrahydrosecamine were determined by means of their spectral data. During the isolation of the antimicrobially active alkaloids some inactive alkaloids were also isolated: compactinervine, N (a)-acetylaspidospermidine, O-dimethylaspidospermine, aricine, yohimbine and O-acetylyohimbine.     

The Bowman-Birk inhibitor. Trypsin- and chymotrypsin-inhibitor from soybeans

Four decades of studies on the isolation, characterization, properties, structure, function and possible uses of the Bowman-Birk trypsin- and chymotrypsin-inhibitor from soybeans are reviewed. Starting from Bowman's Acetone Insoluble factor, designated Ai, AA and SBTIAA, the Bowman-Birk inhibitor (BBI) was found to be a protein molecule consisting of a chain of 71 amino acids cross linked by 7 disulfide bonds, with a tendency to self-associate. BBI possesses two independent sites of inhibition, one at Lys 16-Ser 17 against trypsin and the other at Leu 43-Ser 44 against chymotrypsin. It forms a 1:1 complex with either trypsin or chymotrypsin and a ternary complex with both enzymes. Ingestion of BBI by rats, chicks or quails affects the size and protein biosynthesis of the pancreas. Establishment of the full covalent structure of BBI revealed a high homology in the sequences around the two inhibitory sites, suggesting evolutionary gene duplication from a single-headed ancestral inhibitor. Scission of BBI by CNBr followed by pepsin results in two active fragments, one that inhibits trypsin and the other, chymotrypsin. Replacements and substitutions in the reactive sites result in changes in inhibitory activity and in specificity of inhibition. Conformation studies, labeling of BBI with a photoreactive reagent, chemical synthesis of cyclic peptides that include inhibitory sites, in vitro synthesis of BBI, and species specificity regarding the inhibited enzymes are described. The significance of BBI as a prototype of a family of inhibitors present in all legume seeds is discussed.     

榅桲果实及种子不同提取部位的蛋白酪氨酸磷酸酶1B抑制活性筛选

目的探讨榅桲果实和种子的甲醇提取物及其不同溶剂提取部位对蛋白酪氨酸磷酸酶1B(PTP1B)的抑制作用,筛选抗糖尿病活性部位。方法利用PTP1B体外筛选系统,对榅桲果实和种子不同提取部位进行细致筛选,确定其降糖活性部位。结果榅桲果实和种子各部位均有不同程度抑制PTP1B活性的作用,其中榅桲种子乙酸乙酯部位、榅桲果实二氯甲烷部位和榅桲果实水部位的体积分数为50%的乙醇溶液洗脱部位对PTP1B抑制作用强,其半数抑制质量浓度(IC50)分别为0.844,0.423和0.395mg·L-1,初步推测榅桲果实及种子提取物具有抗2型糖尿病的潜力。结论榅桲种子乙酸乙酯部位、榅桲果实二氯甲烷部位和榅桲果实水部位的体积分数为50%的乙醇溶液洗脱部位是主要的抗糖尿病活性部位。     

川芎嗪抗肿瘤转移作用及其机理

川芎嗪(TTMP)在20mg·kg~(-1)·d~(-1)剂量下,给药18d,能显著抑制B16-F10黑素瘤的人工肺转移,其肺转移结节数由134个下降到72个.放射免疫法测定TTMP能显著降低肺转移小鼠血浆TXB_2含量,而对6-keto-PGF_(1α)含量无显著影响.同位素参入法测定TTMP能增强正常及荷瘤小鼠脾脏NK细胞活性,且能拮抗环磷酰胺对NK细胞活性的抑制作用.TTMP抗肿瘤转移作用可能与降低小鼠血浆TXB_2含量和增强NK细胞活性有关     

Antifungal activity of geraniol and citronellol,two monoterpenes alcohols,against Trichophyton rubrum involves inhibition of ergosterol biosynthesis

Abstract

Context: Trichophyton rubrum is the most common fungus causing chronic dermatophytosis in humans. Antifungal activity of promising agents is of great interest. Geraniol and citronellol are monoterpenes with antimicrobial properties.

Objective: This study aimed to investigate the inhibitory effects and possible mechanism of antifungal activity of geraniol and citronellol against strains of T. rubrum.

Materials and methods: The minimum inhibitory concentration (MIC) of each drug against 14 strains was determined by broth microdilution. The effects of the drugs on dry mycelial weight, conidial germination, infectivity on human nail fragments, and morphogenesis of T. rubrum were analyzed. The effects on the cell wall (test with sorbitol) and cell membrane (release of intracellular material and ergosterol biosynthesis) were investigated.

Results: MIC values of geraniol ranged between 16 and 256?µg/mL while citronellol showed MIC values from 8 to 1024?µg/mL. The drugs (MIC and 2?×?MIC) inhibited the mycelial growth, conidia germination, and fungal growth on nail fragments. The drugs (half of MIC) induced the formation of wide, short, and crooked hyphae in T. rubrum morphology. With sorbitol, geraniol MIC was increased by 64-fold and citronellol by 32-fold. The drugs caused leakage of intracellular material and inhibited ergosterol biosynthesis.

Discussion: The results suggest that the drugs damage cell wall and cell membrane of T. rubrum through a mechanism that seems to involve the inhibition of the ergosterol biosynthesis.

Conclusion: This study confirms that geraniol and citronellol can be regarded as potential drugs for controlling T. rubrum growth, with great potential against agents of dermatophytosis.     

苯烷基异硫脲类化合物的合成及其一氧化氮合酶抑制活性苯烷基异硫脲类化合物的合成及其一氧化氮合酶抑制活性

目的寻找抑制神经型一氧化氮合酶(nNOS)并对痴呆症等有较好治疗作用的化合物。方法以异硫脲为母核,在其结构中引入苯烷基得到目的物,通过药物对神经细胞中亚硝酸盐含量的影响，测定目的物对nNOS的抑制作用。结果设计合成了16个新的苯烷基异硫脲类化合物(I1～16)。其结构经MS,IR,¹HNMR和元素分析确证。所有目的物均有不同程度的nNOS抑制活性，其中化合物I₈,I₁₂和I₁₄的活性较强。结论化合物I₈,I₁₂和I₁₄的活性强于阳性对照药S-甲基-N-(4-甲氧基苯基)异硫脲,IC₅₀达到1×10-7 mol·L^-1水平。