多荚草皂甙D和E的结构

从石竹科植物多荚草（Polycarpon prostratum(Forssk)Aschers.et Schwein.ex Aschers）的全草中分离得到了2个新的柴胡皂甙类化合物：prostratoside D和E。它们的结构过波谱方法分别鉴定为3－O－{β－D－xylpoyranosy-(1→2）－β－D－glucopyransoyl-(1→4）－[β－D－glucopyranosyl-（1→2）]-α－L－arabinopyranoside}－saikogeninD和3-O-{β－D－xylpoyranosy-(1→2）－β－D－glucopyransoyl-(1→4）－[β－D－glucopyranosyl-（1→2）]-α－L－arabinopyranoside}－saikogeninG。     

金边瑞香有机相提取物库的构建及活性成分分离纯化研究

采用逆流提取-系统溶剂组分化-高效液相色谱法(high-performance liquid chromatography,HPLC)检测组分化效果-高速逆流色谱技术(High-Speed Counter-Current Chromatography,HSCCC)分离制备高纯度化合物的研究思路,对金边瑞香化学成分进行研究得到4个高纯度单体化合物.通过现代谱学方法分别鉴定为:瑞香素(daphnetin,1)、瑞香黄烷I(daphnodorin I,2)、对羟基苯甲酸(p-hydroxybenzonic acid,3)和瑞香黄烷D1(daphnodorin D1,4),化合物2～4均为首次从该种植物中分离得到.     

口蹄疫病毒3D蛋白在体外的表达纯化及二级结构分析

口蹄疫(FMD)是严重影响全球农业经济的高度传染的毁灭性的疫病.3D蛋白是口蹄疫病毒(FMDV)编码的RNA聚合酶,参与病毒RNA的复制.利用PCR扩增得到了FMDV的3D基因片段,然后将其克隆到原核表达质粒pET-28a(+)上,构建重组表达质粒pETFM3D.转化宿主菌BL21(DE3)后,利用1 mmol/L的IPTG进行诱导表达.SDS-PAGE和Western boltting分析结果表明,得到了稳定、过量表达的可溶性的3D蛋白质.目的蛋白质经NI亲和层析柱一步纯化就达到95%,再经Q-sepharose柱纯化后达到97%.通过圆二(CD)色谱测定和计算3D蛋白质在不同的pH(2、4、6、8和10)和不同的温度下(25～85℃)的二级结构.上述结果表明,表达的可溶性3D蛋白质具有高表达、易纯化和稳定性好等特点.     

海洋链霉菌Streptomyces sp.SCSIO 1667中吲哚咔唑生物碱类成分的研究

运用大孔树脂柱及硅胶柱层析对南海海洋链霉菌Streptomyces sp.SCSIO 1667的代谢产物进行了研究,从SCSIO 1667菌株的发酵产物中分离得到两个indolocarbazole生物碱类化合物,经质谱,1D、2D NMR波谱数据分析鉴定为星形孢菌素(staurosporine,1)和K-252d(2)。     

放线菌Streptomyces nojiriensis SCSIO m34-1吩嗪生物碱类次级代谢产物的研究北大核心CSCD

从我国黑龙江漠河泥土中分离得到一株放线菌,通过16S rRNA基因测序分析,鉴定该菌株为Streptomyces nojiriensis SCSIO m34-1。采用摇瓶发酵方法对该菌株进行发酵,发酵产物经硅胶柱色谱及半制备高效液相分离得到7个化合物,利用HR-MS、1D和2D NMR波谱数据分析,鉴定其结构分别为izuminoside D(1)、izuminoside A(2)、solphenazine E(3)、solphenazine D(4)、solphenazine A(5)、6-羟基吩嗪-1-羧酸甲酯(methyl 6-hydroxyphenazine-1-carboxylate,6)和diastaphenazine(7),其中izuminoside D(1)是新化合物。抗真菌活性测试结果表明izuminoside D(1)在浓度为50μg/m L时,对苹果腐烂病菌(Valsa mali)和小麦赤霉病菌(Gibberella sanbinetti)有微弱抑制活性。     

一株产虾青素酵母菌株的鉴定

从市售海水鱼内脏中分离得到一株产虾青素的酵母,编号为NZ- 01.采用传统形态学鉴定方法及rDNA序列分析法分别对从NZ- 01进行鉴定.形态学鉴定结果表明该菌为胶红酵母(Rhodotorula mucilaginosa),分别用特异性引物对rDNA 序列的18S rDNA D1/D2区、26S rDNA D1/D2区和ITS区进行扩增,PCR产物测序,将测序结果登录GenBank进行BLAST分析,结果均与形态学观察结果一致,NZ- 01为胶红酵母.     

Ⅰ型鸭肝炎病毒VP1、3D基因克隆及其在大肠杆菌中的表达

根据GenBank中的Ⅰ型鸭肝炎病毒全基因序列设计了扩增Ⅰ型鸭肝炎病毒VP1、3D基因的引物,用该特异性表达引物从Ⅰ型鸭肝炎病毒cDNA模板中扩增得到目的基因VP1、3D,用相同的限制性内切酶酶切目的基因和表达载体pET32a后构建重组表达载体,转化宿主BL21(DE3),用不同浓度的IPTG诱导VP1、3D基因的表达,收集菌液进行SDS-PAGE电泳,Western-blotting分析蛋白免疫原性.结果表明,VP1、3D在大肠杆菌中表达量较高,表达产物的分子量约为48 kD、68 kD,并能被兔抗DHV-1血清所识别.Ⅰ型鸭肝炎病毒VP1、3D蛋白在大肠杆菌中表达产物具有免疫原性.     

放线菌Streptomyces nojiriensis SCSIO m34-1吩嗪生物碱类次级代谢产物的研究

从我国黑龙江漠河泥土中分离得到一株放线菌,通过16S rRNA基因测序分析,鉴定该菌株为Streptomyces nojiriensis SCSIO m34-1。采用摇瓶发酵方法对该菌株进行发酵,发酵产物经硅胶柱色谱及半制备高效液相分离得到7个化合物,利用HR-MS、1D和2D NMR波谱数据分析,鉴定其结构分别为izuminoside D(1)、izuminoside A(2)、solphenazine E(3)、solphenazine D(4)、solphenazine A(5)、6-羟基吩嗪-1-羧酸甲酯(methyl 6-hydroxyphenazine-1-carboxylate,6)和diastaphenazine(7),其中izuminoside D(1)是新化合物。抗真菌活性测试结果表明izuminoside D(1)在浓度为50μg/m L时,对苹果腐烂病菌(Valsa mali)和小麦赤霉病菌(Gibberella sanbinetti)有微弱抑制活性。     

羽叶鬼灯檠中的单萜二糖苷

从羽叶鬼灯檠 (RodgersiapinnataFranch .)的根茎中分离得到 6个单萜二糖苷 ,它们的结构通过波谱方法分别鉴定为 :(E ) 3,7 dimethyl 1 O [α L rhamnopyranosyl (1→ 6 ) β D glu copyranosyl] oct 2 en 7 ol (1) ,(E ) 3,7 dimethyl 1 O [α L arabinofuranosyl (1→ 6 ) β D glucopy ranosyl] oct 2 en 7 ol (2 ) ,geranyl 1 O α L arabinofuranosyl (1→ 6 ) β D glucopyranoside (3) ,gera nyl 1 O α L rhamnopyranosyl (1→ 6 ) β D glucopyranoside (4 ) ,geranyl 1 O β D xylopyranosyl (1→6 ) β D glucopyranoside (5 ) ,geranyl 1 O α L arabinopyranosyl (1→ 6 ) β D glucopyranoside (6 )。其中化合物 1为新化合物 ,单萜二糖苷类化合物系首次在该属中发现。     

魔芋葡甘露寡糖的制备、化学结构及对胰岛NO自由基释放量的影响

魔芋精粉经 β 甘露聚糖酶酶解成寡糖后 ,用活性炭柱进行分离纯化 ,以不同浓度 (5 % ,10 % ,2 0 % )的乙醇洗脱 .研究不同洗脱组分对链脲佐菌素 (STZ)诱导糖尿病模型的胰岛NO自由基释放量的影响 .发现 1mg ml以 5 %乙醇洗脱的寡糖可以使胰岛培养液中的NO自由基释放量平均下降2 5 4 % (P <0 0 5 ) ,0 1mg ml以 5 %乙醇洗脱的寡糖使NO自由基水平下降 2 0 % (P <0 0 5 ) .结果表明 ,5 %乙醇洗脱的魔芋寡糖对保护胰岛免受链脲佐菌素 (STZ)的破坏有一定的作用 .用凝胶色谱、红外光谱、元素分析、核磁共振光谱、质谱等方法初步分析了 5 %乙醇洗脱的魔芋寡糖的化学结构 .发现该糖是一种四糖 ,分子量为 6 6 6 .其推测性结构式为 :β D Man(1→ 4 ) β D Man(1→ 4 ) β D Glc(1→ 4 )α D Man ,β D Man(1→ 4 ) β D Glc(1→ 4 ) β D Man(1→ 4 )α D Man或 β D Glc(1→ 4 ) β D Man(1→4 ) β D Man(1→ 4 )α D Man .     

A-active trisaccharides isolated from A1 and A2 blood-group-specific glycoproteins

Blood-group-specific glycoproteins obtained from ovarian cyst fluids of A1 and A2 persons were degraded with NaOH/NaBH4. The oligosaccharides released were de-N-acetylated with Ba(OH)2 and then hydrolysed with dilute H2SO4. The products were fractionated on columns of ion-exchange resin and the components isolated were re-N-acetylated with 14C-labelled acetic anhydride; further purification was effected by paper chromatography. The following trisaccharides: type 1, GalNAc(alpha 1-3)Gal(beta 1-3)GlcNAc; type 2, GalNAc(alpha 1-3)-Gal(beta 1-4)GlcNAc; type 3 (reduced), GalNAc(alpha 1-3)Gal (beta 1-3)GalNAcOH (where Gal is galactose; GalNAc is N-acetylgalactosamine, GlcNAc is N-acetylglucosamine and GalNAcOH is N-acetylgalactosaminitol) were isolated and characterised from both the A1 and A2 materials. The type 3 (reduced) trisaccharide has not previously been obtained from human glycoproteins. Chromatographic evidence indicated that the three trisaccharide structures were also present in other A1, A2, A1B and A2B ovarian cyst glycoproteins and in A1 and A2 salivary glycoproteins. These findings are not indicative of structural differences between the A determinants of A1 and A2 glycoproteins.     

The protein phosphatases involved in cellular regulation. 2. Purification, subunit structure and properties of protein phosphatases-2A0, 2A1, and 2A2 from rabbit skeletal muscle

Protein phosphatases-2A0, 2A1 and 2A2 have been purified to homogeneity from rabbit skeletal muscle. Approximately 1 mg of phosphatase-2A0 and 2A1, and 0.5 mg of phosphatase-2A2, was isolated from 4000 g muscle within 10 days. Protein phosphatases-2A0 and 2A1 each comprised three subunits, termed A, B' and C (2A0) or A, B and C (2A1), while phosphatase-2A2 contained only two subunits, A and C. The A and C components of phosphatases-2A0, 2A1 and 2A2 had indistinguishable mobilities on sodium dodecyl sulphate/polyacrylamide gels and identical peptide maps. By these criteria, the C component was also identical to the catalytic subunit of phosphatase-2A purified from ethanol-treated muscle extracts. The electrophoretic mobilities of the B and B' subunits were slightly different, and their peptide maps were distinct. The molecular masses of the native enzymes determined by sedimentation equilibrium centrifugation were 181 +/- 6 kDa (2A0), 202 +/- 6 kDa (2A1) and 107 +/- 5 kDa (2A2), while those of the subunits estimated by sodium dodecyl sulphate/polyacrylamide gel electrophoresis were 60 kDa (A), 55 kDa (B), 54 kDa (B') and 36 kDa (C). These values, in conjunction with molar ratios estimated by densitometric analyses of the gels, suggest that the subunit structures of the enzymes are AB'C2 (2A0), ABC2 (2A1) and AC (2A2). Protein phosphatase-2A2 appears to be derived from 2A0 and/or 2A1 during purification through degradation or dissociation of the B' and/or B subunits. Protein phosphatases-2A0, 2A1 and 2A2 were the only phosphorylase phosphatases in rabbit skeletal muscle that were activated by the basic proteins, protamine (A0.5 = 0.25 microM), histone H1 (A0.5 = 0.3 microM) and polylysine (A0.5 = 0.04 microM). Activation by protamine varied over 5-20-fold for phosphatase-2A0 and 5-7-fold for phosphatases-2A1 and 2A2. The dephosphorylation of glycogen synthase was activated by basic proteins in a similar manner to the phosphorylase phosphatase activity. The isolated C subunit was also stimulated by histone H1 and protamine, but 5-10-fold higher concentrations were required, and with phosphorylase as substrate, maximum activation was only about 2-fold. Activation by basic proteins appears to involve their interaction with the A and/or C subunits, but not with the B or B' subunits, or substrates phosphorylase and glycogen synthase.     

Purification and characterization of kallikrein-like serine proteases from rat submandibular glands.

The purification and characterization of kallikrein-like proteases from rat submandibular glands is described. The proteolytic activity of each fraction during purification was monitored on the synthetic substrate N-alpha-tosyl-L-arginine methyl ester (TAME). The purification scheme involved ammonium sulfate precipitation, chromatography on columns of DEAE-Sepharose and Sephadex G-100 and chromatofocusing. Three TAME-hydrolytic activity peaks were eluted from DEAE-Sepharose as unbound fraction (Pool 1), at 125 mM NaCl (Pool 2) and at 250 mM NaCl concentration (Pool 4). Pool 1 further resolved into two protease fractions (1A1 and 1A2), pool 2 into three protease fractions (2A1, 2A2 and 2A3) and pool 4 gave a single major protease peak (4A1) by chromatofocusing on PBE-94. Protease pools 2A2, 2A3, and 4A1 each gave a single band on SDS-polyacrylamide gel electrophoresis with an estimated molecular weight of 34 kDa, 46 kDa and 46 kDa respectively. Pools 1A1, 1A2, 2A1 and 2a2 gave a single precipitin line with anti-rat glandular kallikrein antibodies. 2A3 and 4A1 did not react with these antibodies. Synthetic substrates DL-val-leu-arg-pNA and Bz-pro-phe-arg-pNA, specific for kallikrein-like proteases, were hydrolyzed preferentially by 2A3 and 4A1 but were poor substrates for 1A1, 1A2, 2A1 and 2A2.     

Polymorphisms of alpha-1-acid (orosomucoid), alpha-2-HS-glycoproteins and alpha-1-B among the Parsis of India.

Genetic polymorphisms of plasma alpha 1-acid glycoprotein (oro-somucoid, ORM), alpha 2-HS-glycoprotein (A2HS) and alpha 1-B-glycoprotein (alpha 1B) were studied in a group of Parsis in Bombay, India. The frequencies of ORM1*1, ORM1*2 and ORM1*3 were found to be 0.636, 0.356 and 0.008, respectively. A2HS*1, A2HS*2 and A2HS*3 frequencies were 0.855, 0.135 and 0.010, while the frequencies of A1B*1 and A1B*2 were 0.881 and 0.119, respectively. The phenotype distribution at all three loci was at Hardy-Weinberg equilibrium. The ORM2 locus was monomorphic in the Parsis.     

Genetic Polymorphisms of Sulfotransferases (SULT1A1 and SULT1A2) in a Turkish Population

Sulfotransferases (SULTs) play a significant role in the biotransformation of a variety of xenobiotics and endogenous compounds. SULTs are genetically polymorphic enzymes; to date, 12 human cytosolic SULT isoforms have been identified. This study investigated SULT1A1 and SULT1A2 gene polymorphism using a PCR-RFLP method (n = 303). The frequency of the SULT1A1*1 allele was 76.2% and SULT1A1*2 was 23.8%. The SULT1A1*3 allele could not be identified. The SULT1A2 frequencies were 69.2% (SULT1A2*1), 18.3% (SULT1A2*2), and 12.5% (SULT1A2*3). The SULT1A1 and SULT1A2 loci were in Hardy-Weinberg equilibrium (SULT1A1 χ2 = 0.58, P = 0.44; SULT1A2 χ2 = 7.28, P = 0.06). Linkage analysis indicated a close linkage between these two genes (χ2 = 5.31, P < 0.01); therefore, the statistical hypothesis that SULT1A1 and SULT1A2 alleles are independently distributed was rejected. Additionally, a strongly positive linkage was detected between SULT1A1*2 and SULT1A2*2 alleles in this population (D' = 0.79, χ2 = 33.33).     

Concept formation based on the relations among values of multi-component figures

A complex discrimination procedure was used to test class formation with multi-component figures in college students. First, selections of a red-+45 degrees -oriented rectangle (A1B1) instead of a red -45 degrees -oriented rectangle (A1B2) and of a green -45 degrees -oriented rectangle (A2B2) instead of a green-+45 degrees -oriented rectangle (A2B1), were reinforced. Second, selections of a +45 degrees -obtuse-angle-white hexagon (B1C1), instead of a +45 degrees -two-acute-angle hexagon (B1C2) and of a -45 degrees -two-acute-angle hexagon (B2C2), instead of a -45 degrees -obtuse-angle hexagon (B2C1), were reinforced. Subsequent tests with figures A1B1 and A2B1, A2B2 and A1B2, B1C1 and B2C1, and B2C2 and B1C2 demonstrated selection of the same figures as in training. Other tests with a novel figure compounded by color A1 and form C1 and a novel figure compounded by color A1 and form C2, or a novel figure compounded by A2 and C1 and a novel figure compounded by A2 and C2 demonstrated consistent selection of figures A1C1 and A2C2. A final test with figures A1C1 and A2C1, or with figures A2C2 and A1C2 also showed consistent selections of the same figures -A1C1 and A2C2. The resulting classification among selected and non-selected figures cannot be based on particular properties of the selected figures, or in a combination of a particular set of those properties, because both selected and non-selected figures were compounded by identical values. Instead, the participants selected the figures with two values of the same set (A1,B1, or C1, or A2,B2 or C2). Thus, these results demonstrated categorization based on the relations among the values of the figures. Therefore, these results have important implications for the study of concept formation.     

Frequencies of four genetic polymorphisms in the CYP1A2 gene in Turkish population

Cytochrome P450 (CYP) 1A2 gene is involved in the metabolic activation of several carcinogens and altered metabolization of some clinically used drugs. We aimed to investigate the distributions of genetic polymorphisms -3860 (G/A)(CYP1A2*1C) and -2467 (T/del)(CYP1A2*1D) in the 5'-flanking region and -739 (T/G)(CYP1A2*1E) and -163(C/A)(CYP1A2*1F) in the first intron of the CYP1A2 gene in 110 unrelated healthy Turkish volunteers by PCR-RFLP technique. The frequencies of each polymorphism in Turkish population were found as 0.04, 0.92, 0.01, 0.27 for CYP1A2*1C, CYP1A2*1D, CYP1A2*1E, CYP1A2*1F, respectively. Compared with other populations, CYP1A2*1D has been found to be significantly increased in Turkish population. On the other hand, in general, the frequencies of the other polymorphisms were concordant with those in the Egyptian and Caucasian populations, and were different from those in the Japanese, Chinese and Ethiopian populations. Our results suggest that due to increased frequency of CYP1A2*1D in Turkish population, functional significance of CYP1A2*1D should be evaluated. It might be screened to determine the relationship between CYP1A2*1D and CYP1A2 related drug metabolisms in associated groups.     

Allele and genotype frequencies of the polymorphic cytochrome P450 genes (CYP1A1, CYP3A4, CYP3A5, CYP2C9 and CYP2C19) in the Jordanian population

Drug metabolizing enzymes participate in the neutralizing of xenobiotics and biotransformation of drugs. Human cytochrome P450, particularly CYP1A1, CYP2C9, CYP2C19, CYP3A4 and CYP3A5, play an important role in drug metabolism. The genes encoding the CYP enzymes are polymorphic, and extensive data have shown that certain alleles confer reduced enzymatic function. The goal of this study was to determine the frequencies of important allelic variants of CYP1A1, CYP2C9, CYP2C19, CYP3A4 and CYP3A5 in the Jordanian population and compare them with the frequency in other ethnic groups. Genotyping of CYP1A1(m1 and m2), CYP2C9 (*2 and *3), CYP2C19 (*2 and *3), CYP3A4*5, CYP3A5 (*3 and *6), was carried out on Jordanian subjects. Different variants allele were determined using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). CYP1A1 allele frequencies in 290 subjects were 0.764 for CYP1A1*1, 0.165 for CYP1A1*2A and 0.071 for CYP1A1*2C. CYP2C9 allele frequencies in 263 subjects were 0.797 for CYP2C9*1, 0.135 for CYP2C9*2 and 0.068 for CYP2C9*3. For CYP2C19, the frequencies of the wild type (CYP2C19*1) and the nonfunctional (*2 and *3) alleles were 0.877, 0.123 and 0, respectively. Five subjects (3.16?%) were homozygous for *2/*2. Regarding CYP3A4*1B, only 12 subjects out of 173 subjects (6.9?%) were heterozygote with none were mutant for this polymorphism. With respect to CYP3A5, 229 were analyzed, frequencies of CYP3A5*1,*3 and *6 were 0.071, 0.925 and 0.0022, respectively. Comparing our data with that obtained in several Caucasian, African-American and Asian populations, Jordanians are most similar to Caucasians with regard to allelic frequencies of the tested variants of CYP1A1, CYP2C9, CYP2C19, CYP3A4 and CYP3A5.     

不同术前皮肤准备方案与手术切口感染的关系研究

目的:研究不同术前皮肤准备方案与手术切口感染(SSI)的关系,为降低临床SSI发生率提供参考。方法:选择自2015年1月～2019年12月在医院行手术治疗的患者1810例为本次研究对象。根据析因设计表,将因素A:是否剃毛(1不剃毛;2剃毛),B:清洁方式(1清水清洁;2肥皂水清洁),C:术前备皮时间(1术前1 d;2术前2 h)配对分为8个组:A1B1C1组226例,A1B2C1组229例,A1B1C2组216例,A1B2C2组232例,A2B1C1组221例,A2B2C1组241例,A2B1C2组221例,A2B2C2组224例,比较各组手术部位及切口类型分布、术后SSI发生率,并采用析因分析法分析术前皮肤准备后各组菌落计数的相关性及交互作用。结果:各组患者的手术部位及切口类型之间的差异不存在统计学意义(P0.05)。A1B1C1组及A2B1C1组的SSI发生率较高,分别为12.83%和14.48%。A1水平的SSI发生率是8.75%,与A2水平的8.27%相比,差异不存在统计学意义(P0.05)。B1、C1水平的SSI发生率分别是11.31%、10.03%,明显高于B2、C2水平的5.83%、6.94%,差异均存在统计学意义(P0.05)。各组术前皮肤准备后的菌落计数差异存在统计学意义(P0.05),析因分析结果显示,B、C单因素分析差异存在统计学意义(P0.05),且A与C,B与C间具有交互作用,而A、B、C间具有二级交互作用(P0.05)。结论:术前皮肤准备对降低SSI发生具有重要作用,实际操作时,建议在较短的时间内利用肥皂水或其他消毒水进行皮肤清洗并完成备皮。