人OCTN2原核表达载体PGEX-2T-OCTN2的构建及意义

目的选取并克隆人特异性OCTN2基因片段,构建原核表达载体PGEX-2T-OCTN2。方法应用RT-PCR方法从人附睾组织中克隆特异性人OCTN2基因片段,通过双酶切方法把OCTN2基因片段克隆到PGEX-2T载体,再经双酶切和测序鉴定。结果获取并克隆特异性人OCTN2的基因片段,构建原核表达载体PGEX-2T-OCTN2。结论成功构建原核表达载体PGEX-2T-OCTN2,为进一步研究附睾肉碱转运机制奠定前期基础。     

C-erbB-2在乳腺癌中的表达及意义

目的探讨C-erbB-2的表达与乳腺癌病理分级,临床分期的关系。方法应用免疫组织化学方法检测116例乳腺癌中C-erbB-2的表达。结果116例中C-erbB-2的阳性表达率为59.5%。C-erbB-2在病理分级Ⅰ、Ⅱ、Ⅲ级肿瘤中的阳性表达率分别为38.9%、52.9%、80.4%,三组间比较差别有高度显著性(P<0.01);C-erB-2在临床分期Ⅰ-Ⅱ期和Ⅲ-Ⅳ期组的阳性表达率分别为39.6%、73.5%,两组间比较差别有高度显著性(P<0.01)。结论C-erbB-2的表达与乳腺癌恶性程度有关,可以作为检测乳腺癌分期、分级的重要指标。     

EphB2和EphrinB2及EphrinA1在喉癌组织中的表达及意义

目的探讨EphB2、EphrinB2和EphrinA1与喉癌发生发展的关系及探讨喉癌的发病机制。方法应用流式细胞术检测喉癌组织、癌旁组织及喉部正常黏膜组织的EphB2和EphrinA1、EphrinB2蛋白表达的含量。基因蛋白以荧光指数（FI）〉1.0为阳性表达,FI≤1.0为阴性表达。结果EphB2蛋白、EphrinB2蛋白、EphrinA1蛋白在喉癌组织中的阳性表达率分别高于癌旁组织和正常喉黏膜组织（P〈0.05或〈0.01）。在喉癌组织中,EphB2蛋白、EphrinB2蛋白、EphrinA1蛋白在淋巴结转移组的阳性表达率分别高于无淋巴结转移组（P〈0.05）;临床Ⅲ、Ⅳ期组的阳性表达率分别高于临床Ⅰ、Ⅱ期组（P〈0.05或〈0.01）。喉癌组织中EphB2蛋白与EphrinA1蛋白、EphrinB2蛋白有多元线性回归关系,且均为正相关（P〈0.01）。结论EphB2蛋白和EphrinA1、EphrinB2蛋白高表达可能与喉癌的发生、发展有关。     

erbB-2/HER-2在肺癌组织中的异常表达及意义

目的 研究erbB-2／HER-2基因在肺癌组织中的异常表达的意义。方法 应用免疫组织化学的方法检测50例肺癌和20例正常肺组织HER-2(人体第二表皮生长因子受体，Human epidermal-growth—factor receptor2，HER2)蛋白的表达。结果 HER-2／neu在正常肺组织中无过表达，肺癌组织中存在HER-2／neu的过表达，过表达率为66％，肺癌HER-2／neu的过表达更多见于中晚期肺癌，特别是肺腺癌，且与患者术后无瘤三年生存率呈负相关。结论 HER-2／neu可作为评价肺癌(特别是肺腺癌)患者预后的可靠指标。     

Bcl-2在膀胱癌中的表达及意义

目的：探讨Bcl-2蛋白在膀胱移行细胞癌中的分布与生物学行为的关系。方法：应用ABC免疫组织化学法对66例人体膀胱移行细胞癌进行研究。结果：正常膀胱粘膜Bcl-2均为阴性。Bcl-2在浸润性膀胱癌中的阳性表达高于浅表性膀胱癌,但相互间差异不具有统计学意义（P＞0．05）;Bcl-2随病理分级增高阳性表达逐渐提高,相互间差异具有统计学意义（P＜0．05）;在复发肿瘤中的阳性表达高于未复发肿瘤,相互间差异具有统计学意义（P＜0．05）。结论：Bcl-2蛋白在膀胱移行细胞癌中的表达与临床分期无关,与病理分级有关,可作为术后预测肿瘤复发的参考指标之一。     

EphA2在膀胱癌中的表达及意义

目的:探讨EphA2在膀胱癌中的表达及意义。方法:通过免疫组织化学法和RT-PCR法检测EphA2在膀胱癌和正常膀胱组织中的表达。结果:利用免疫组织化学法检测发现EphA2在膀胱癌组织中异常表达率为77.46%(55/71),正常膀胱组织异常表达率为20%(2/10),在膀胱癌组织中EphA2的异常表达率明显高于正常膀胱组织的异常表达率。利用RT-PCR法在基因水平检测膀胱癌组织与正常膀胱组织中EphA2的表达发现:EphA2在膀胱癌组织所检测的光密度值为1.156±0.179,而在正常膀胱组织所得光密度值为0.978±0.086。通过统计学分析,EphA2在膀胱癌组织中的表达与正常膀胱组织相比差异显著(P<0.05)。结论:蛋白和基因水平的同时检测EphA2在膀胱癌组织中的异常表达率都高于正常膀胱组织。     

COX-2在乳腺癌组织中的表达及临床意义

目的　研究环氧化酶 2在乳腺癌组织和癌旁组织中的表达情况与临床病理指标之间的相关性。方法　以免疫组化SP法检测 4 7例乳腺癌以及癌旁组织标本。结果　 4 7例癌组织标本中阳性率为5 3 2 % (2 5 / 4 7)显著高于癌旁组织 (6 38% ,3/ 4 7) ,在淋巴结转移阳性组和阴性组以及不同组织学分级的病人之间COX 2的表达差异有显著意义 (P <0 0 5 ) ,而与病人年龄、月经情况、肿瘤大小、临床分期、ER、PR等差异无显著意义 (P >0 0 5 )。结论　乳腺癌组织中存在环氧化酶 2的高表达 (5 3 2 % ) ,并与淋巴结转移和组织学分级有关 ,环氧化酶 2在乳腺癌的发生发展中可能扮演重要角色     

乳腺癌COX-2、bFGF的表达及临床意义

目的：研究环氧化酶-2（Cyclooxygenase-2,COX-2）碱性成纤维细胞生长因子（basic fibrob last growth factor,bFGF）在乳腺癌中的表达及意义。方法：应用S-P免疫组化法检测38例乳腺癌和20例乳腺正常组织中COX-2、bFGF的表达,使用统计学方法分析上述因子表达与乳腺癌肿瘤直径、临床分期、病理分级、淋巴转移之间的关系。结果：COX-2、bFGF在乳腺癌中高表达,在正常乳腺表达水平较低,差异有统计学意义（P〈0.05）;COX-2、bFGF的表达与肿瘤体积、临床分期、病理分级、淋巴结转移情况关系密切,与肿瘤的发病年龄无明显相关性。结论：通过抑制COX-2和bFGF的高表达来抑制乳腺癌的发生、发展、增殖、转移可能会成为一项有效的乳腺癌辅助治疗措施。     

β-内酰胺类抗生素过敏患者血清IL-2及sIL-2R表达

目的 探讨白细胞介素2(IL-2)及可溶性白细胞介素2受体(sIL-2R)与β-内酰胺类药物过敏之间的关系.方法 采用ELISA检测70例β-内酰胺类药物过敏反应患者(A组)与20例正常人(B组)外周血血清IL-2及sIL-2R的水平.采用放射免疫吸附(RAST)方法检测过敏患者血清中特异性IgE抗体.结果 A组血清IL-2水平显著高于B组(P<0.05)(4.110 ng/ml vs.2.070ng/ml).A组中IgE抗体阳性患者sIL-2R水平高于抗体阴性者(3.300 ng/ml vs.2.345 ng/ml)(P<0.05).结论 β-内酰胺类抗生素过敏反应的发生中,外周血血清IL-2水平升高;特异性IgE抗体阳性的过敏反应与sIL-2R水平升高有关.     

SSTR2在血管瘤中的表达及统计分析

目的:应用免疫组织化学方法检测不同时期血管瘤组织生长抑素受体2(Somatostatin receptor 2,SSTR2)的表达,探讨其在血管瘤发生、发展过程中的作用机制。方法:收集武汉大学人民医院病理科2005年9月～2009年12月皮肤毛细血管瘤存档蜡块50例,其中男性25例,女性25例。采用免疫组织化学S-P法检测50例皮肤血管瘤增生期、退化期及正常皮肤组织SSTR2的表达水平,采用HPIAS-1000图文报告管理系统对SSTR2的表达进行定量分析,并用SPSS11.5软件对各组免疫组织化学反应阳性颗粒的平均光密度、阳性面积率做单因素方差分析和SNK(q)检验。结果:增生期血管瘤血管内皮细胞中可见少量的棕黄色颗粒,SSTR2低表达,正常皮肤组及退化组血管内皮细胞细胞中可见密集分布的棕黄色颗粒,SSTR2呈高表达。增生期组SSTR2的表达明显低于退化期组和正常皮肤组(P<0.05),而后两组比较差异无统计学意义(P>0.05)。结论:SSTR2在血管瘤的发生、发展中起了重要作用。     

2-吡啶甲醇及2-吡啶甲醛的合成

以 2 -甲基吡啶为原料、过氧化氢为氧化剂制备 2 -吡啶甲醇和 2 -吡啶甲醛,工艺方法经济、安全     

Synthesis and immunosuppressive activity of 2-substituted 2-aminopropane-1,3-diols and 2-aminoethanols

A series of 2-substituted 2-aminopropane-1,3-diols was synthesized and evaluated for their lymphocyte-decreasing effect and immunosuppressive effect on rat skin allograft. A phenyl ring was introduced into the alkyl chain of the lead compound 3, which is an immunosuppressive agent structurally simplified from myriocin (1, ISP-I) via compound 2. The potency of the various compounds was dependent upon the position of the phenyl ring within the alkyl side chain. The most suitable length between the quaternary carbon atom and the phenyl ring was two carbon atoms. 2-Substituted 2-aminoethanols were successively synthesized and evaluated for their T-cell-decreasing effect and immunosuppressive effect using a popliteal lymph node gain assay in rats. The absolute configuration at the quaternary carbon affected the activity, and the (pro-S)-hydroxymethyl group of compound 6 was essential for potent immunosuppressive activity. Favorable substituents for the (pro-R)-hydroxymethyl group of 6 were hydroxyalkyl (hydroxyethyl and hydroxypropyl) or lower alkyl (methyl and ethyl) groups. 2-Amino-2-[2-(4-octylphenyl)ethyl]propane-1,3-diol hydrochloride (6, FTY720) was found to possess considerable activity and is expected to be useful as an immunosuppressive drug for organ transplantation.     

2-苯基-1-丁醇与2-二甲氨基-2-苯基丁腈的合成

目的为马来酸曲美布汀的重要中间体2-二甲氨基-2-苯基-1-丁醇的合成奠定基础。方法苯乙腈与溴乙烷进行烃化反应得2-苯基-1-丁腈,所得产物经水解得2-苯基-1-丁酸,然后通过硼氢化钠-碘体系还原得2-苯基-1-丁醇;苯乙腈与N-溴代丁二酰亚胺进行卤代反应得溴代苯乙腈,所得产物与二甲胺进行烃化反应得2-二甲氨基苯乙腈,然后与溴乙烷进行烃化反应得2-二甲氨基-2-苯基丁腈。结果合成了2-苯基-1-丁醇和2-二甲氨基-2-苯基丁腈,总收率为分别为51%和59.4%。目标产物的结构经核磁共振氢谱、质谱确证。结论本合成方法原料易得,操作简单,收率较高,适合于工业化生产。     

Molecular modeling of 2-alkyloxy- and 2-aralkyloxy-adenosine A1- and A2-agonists

The C2-region of adenosine A1- and A2-receptors by a molecular modeling technique has been extended and applied to a series of 2-substituted adenosines reported by Olsson, et al. The similarity and dissimilarity of the structure maps obtained by molecular modeling have been used as a basis for the mapping of the analysed receptor domain. The proposed model of the C2-region of the A1-receptor consists of a narrow and sterically limited area that interacts well electrostatically with small and electron rich moieties. Olsson's provisional model of the C2-region of the A2-receptor has been extended with two subsites, as well as with a forbidden area near the C2-position of the purine ring. The conformational analysis performed in the study does not support the hypothesis of Olsson et al. that adenosine C2 substituents may partly occupy the same receptor domain as the N6 substituents of the A1-receptor. The occupation of the cycloalkyl subsite increases the receptor selectivity while the occupation of the other subsite by aryl rings, fixed at a parallel position to the purine system, highly enhances the receptor affinity.     

Synthesis and antiviral evaluation of carbocyclic analogues of 2'-azido- and 2'-amino-2'-deoxycytidine

Carbocyclic analogues of 2'-azido- and 2'-amino-2'-deoxycytidine, compounds 8 and 9, were synthesized by an eight-step synthesis from (+/-)-(1 alpha,2 alpha,3 beta,5 beta)-3-amino-5-(hydroxymethyl)-1,2- cyclopentanediol (1), which was prepared from cyclopentadiene via an eight-step route. These compounds were tested in vitro against herpes simplex virus type 1 (HSV-1). The 2'-amino analogue was found to show moderate antiviral activity, with an ED50 of 50 microM. However, the 2'-azido analogue was not active at a concentration up to 400 microM.     

Metabolism and metabolic effects of 2-azahypoxanthine and 2-azaadenosine

The metabolism and metabolic effects of 2-azahypoxanthine and 2-azaadenosine were studied to elucidate the biochemical basis for their known cytotoxicities. 2-Azaadenosine is a known substrate for adenosine kinase. That 2-azahypoxanthine is a substrate for hypoxanthine (guanine) phosphoribosyltransferase is shown by the observations that, in cell-free fractions from HEp-2 cells supplemented with 5-phosphoribosyl-1-pyrophosphate, 2-azahypoxanthine inhibited the conversion of hypoxanthine to IMP but not the conversion of adenine to AMP, and hypoxanthine, but not adenine, inhibited the conversion of 2-azahypoxanthine to 2-azaIMP. [8-14C]2-Azahypoxanthine was synthesized from [8-14C]hypoxanthine via [2-14C]-4-amino-5-imidazolecarboxamide. In HEp-2 cells in culture, the principal metabolite of [8-14C]-2-azahypoxanthine was 2-azaATP; there was no detectable 14C in deoxynucleotides or in DNA or RNA fractions. 2-Azaadenosine was much more toxic than 2-azahypoxanthine, and, when used in the presence of an adenosine deaminase inhibitor, 2'-deoxycoformycin, was converted in HEp-2 cells to 2-azaATP in amounts that exceeded those of ATP in control cells. The pool of ATP was reduced by as much as 75% as 2-azaATP accumulated. In a short-term experiment (4 hr), 2-azaadenosine selectively reduced the pools of adenine nucleotides, whereas 2-azahypoxanthine reduced the pools of guanine nucleotides selectively. Both 2-azahypoxanthine and 2-azaadenosine inhibited the incorporation of formate into purine nucleotides and were without effect on the conversion of thymidine and uridine to nucleotides. 2-Azahypoxanthine inhibited the incorporation of thymidine into macro-molecules but not that of uridine or leucine; 2-azaadenosine inhibited the incorporation of all three of these precursors non-selectively. 2-AzaIMP inhibited IMP dehydrogenase competitively with IMP (Ki = 66 microM). The difference in effects of 2-azahypoxanthine and 2-azaadenosine perhaps may be due to the production, from 2-azahypoxanthine but not from 2-azaadenosine + 2'-deoxycoformycin, of 2-azaIMP, which inhibits synthesis of guanine nucleotides and thereby results in inhibition of DNA synthesis. Specific sites of action for 2-azaadenosine are yet undefined.     

2-Fluoroformycin and 2-aminoformycin. Synthesis and biological activity

Syntheses of 2-fluoroformycin [7-amino-5-fluoro-3-(beta-D-ribofuranosyl)pyrazolo[4,3-d]pyrimidine] (2b) and 2-aminoformycin [5,7-diamino-3-(beta-D-ribofuranosyl)pyrazolo[4,3-d]pyrimidine] (2c) are described. Cytotoxicity data are given for 2b and 2c alone as well as with added pentostatin. Kinetic parameters for adenosine deaminase are also provided. 2-Fluoroformycin, although a much poorer substrate for adenosine deaminase than formycin A, is not nearly as cytotoxic to cells in culture.