双波长紫外分光光度法测定贝母中腺苷和胸苷的含量

将4种贝母的甲醇提取物经薄层色谱法粗分离后,直接用双波长紫外分光光度法测定其含量。结果表明,该方法线性关系好,腺苷和胸苷标准曲线的相关系数均为0.9999,同时也发现平贝、炉贝和伊贝中腺苷都占核苷总量60%以上,而浙贝中仅占约40%,提示贝母生药的抗凝血活性可能与贝母中核苷类化合物的种类和含量的差别有关。     

18F-FDG与18F-FLT PET/CT延迟显像对肺结节诊断效能的评价

目的 通过对多中心、前瞻性研究中接受了18F-脱氧葡萄糖(FDG)与18F-脱氧胸腺嘧啶核苷(FLT)延迟显像病例的分析,探讨18F-FDG与18F-FLT延迟显像对肺结节诊断的效能.方法 6个PET/CT中心,从2006年1月至2007年6月,按照统一标准,采用同机型、同一扫描条件,开展了肺结节样病变18F-FLT和18F-FDG PET/CT显像的多中心临床研究.在经确诊的55例病例中,25例患者进行了18F-FLT显像和延迟显像,34例患者进行了18F-FDG延迟显像.按常规计算延迟显像时病灶最大标准摄取值(SUVmax)及与早期显像时SUVmax相比的变化率(△SUVmax).对照临床确诊结果分析其诊断效能.采用SPSS11.0软件进行统计学处理.结果 18F-FDG延迟显像患者中,6例肺癌中5例、12例结核中9例、16例炎症或其他良性结节中9例的SUVmax较早期相升高.18F-FLT延迟显像组中,7例肺癌中3例、8例结核中3例和10例其他良性病灶中2例的SUVmax上升.经分组统计分析,不同疾病组间18F-FDG延迟显像SUVmax和△SUVmax差异无统计学意义;18F-FLT延迟显像SUVmax和△SUVmax组间差异也无统计学意义.无论18F-FDG还是18F-FLT,延迟显像的诊断效能均不如早期相.无论早期还是延迟显像,单独18F-FDG或18F-FLT显像的诊断效能均不如二者联合应用.结论 18F-FDG和18F-FLT延迟显像的SUVmax变化规律性不强,不宜单独应用于肺结节的鉴别诊断.     

TS、TP及DPD表达与乳腺癌新辅助化疗的关系

目的 检测乳腺癌标本中胸苷酸合成酶（Thymidylate Synthase,TS）、胸腺嘧啶脱氧核苷磷酸化酶（Thymidine Phosphorylase,TP）、二氟嘧啶脱氢酶（Dihydropyrimidine Dehydrogenase,DPD）表达状况,评价TS、TP、DPD的表达与新辅助化疗效果之间的关系.方法 选择81例可手术的乳腺癌患者,手术前均接受CMF方案新辅助化疗,采用免疫组织化学染色的方法对新辅助化疗前麦默通活检系统切取的乳腺癌组织中TS、TP和DPD的表达进行检测.结果 TS、TP阴性患者的部分缓解率要明显高于阳性患者,而DPD阳性患者的部分缓解率明显高于阴性患者,差异具有统计学意义（P＜0.05）;DPD阴性患者Ⅲ～Ⅳ度毒副反应发生率高于阳性患者（P＜0.05）,而TS、TP的表达水平与化疗所产生的毒副反应之间无明显关系（P＞0.05）.结论 TS、TP及DPD的表达与乳腺癌患者接受CMF方案的新辅助化疗的治疗效果相关,而DPD的表达则与含5-FU类化疗药物所产生的毒副反应相关.     

Capacitively coupled electrical stimulation of bovine growth plate chondrocytes grown in pellet form

Pellets formed from isolated bovine growth plate chondrocytes were grown in various capacitively coupled electrical fields. The signals chosen were 0, 10, 100, 250, 500, 750, 1,000, and 1,500 V peak-to-peak, 60 kHz. The effect on cell proliferation and matrix production of these different voltages was determined by [3H]thymidine and [35S]sulfate uptake, respectively, Cyclic AMP assays were done to determine if increases in either thymidine or sulfate uptake were associated with changes in cAMP levels. Significantly increased cell proliferation occurred at 500, 750, and 1,000 V peak to peak. The calculated electric fields were 1.5 to 3.0 x 10(-2) V/cm. Proliferation was significantly inhibited at 1,500 V peak-to-peak with a calculated field of 4.5 x 10(-2) V/cm. Little if any change was seen in cAMP levels at 30 or 60 min following application of the appropriate electric signals.     

Effect of nitric oxide on mitogenesis and proliferation of cerebellar glial cells

In the brain, nitric oxide (NO) has been identified as a messenger molecule and a mediator of excitatory amino acid-induced neurotoxicity. In this study, the effects of NO on serum-induced mitogenesis and cell proliferation of the cerebellar glial cells were assessed. NO-generating agent, S-nitroso-N-acetylpenicillamine (SNAP) increased intracellular cyclic guanosine monophosphate (cGMP) levels. Furthermore, 2 chemically dissimilar NO-generating agents, SNAP and sodium nitroprusside (SNP) inhibited serum-induced thymidine incorporation and cell proliferation. The antimitogenic effect of NO was mimicked by 8-bromo-cGMP and blocked by hemoglobin, a known inhibitor of NO. The effect of NO was not cytotoxic, since the cells were not stained with Trypan blue and did not show increased release of lactate dehydrogenase in the culture supernatants. However, NO-treated cells showed decreased conversion of tetrazolium to blue formazan suggesting that NO inhibited mitochondrial activity in the glial cells. These results demonstrate that NO inhibits serum-induced mitogenesis and cell proliferation of cultured rat cerebellar glial cells.     

原发性食管癌中胸苷磷酸化酶表达和血管生成的临床意义

目的　观察原发性食管癌中胸苷磷酸化酶 (thymidinephosphorylase,TP)的表达情况 ,探讨肿瘤组织中胸苷磷酸化酶表达、肿瘤微血管密度 (microvesseldensity ,MVD)和临床病理特征之间的关系 ,分析TP表达和肿瘤MVD的预后意义。方法　应用单克隆抗体对 6 5例食管癌标本进行免疫组化染色 ,测定TP表达及MVD。结果　TP在食管癌中的表达 (4 5 / 6 5 ,6 9.2 % )明显 (P <0 .0 0 1)高于正常食管粘膜 (4 / 2 4 ,16 .7% )。食管癌的MVD(4 5 .0 9± 8.76 )与正常食管粘膜的MVD(2 7.4 8± 8.4 4 )的差别显著 (P <0 .0 0 1)。食管癌TP阳性的MVD的均值是 (4 6 .5 3± 7.18) ,TP阴性的MVD的均值是 (4 1.85± 11.0 8) ,前者明显 (P =0 .0 4 6 ) 高于后者。食管癌的TP表达与临床病理特征无相关性 ,MVD却与肿瘤的浸润深度(P =0 .0 35 ) 及分期(P =0 .0 18) 有关 ,而且只有MVD才是食管癌的一个预后指标(P <0 .0 0 1)。结论　食管癌的TP表达与MVD密切相关。TP表达与食管癌的临床病理特征无关。MVD与肿瘤的浸润发展有关 ,同时只有MVD才是食管癌的一个预后指标     

Gene transfer by viral vectors for gene therapy

Conclusions The search for useful virus vectors and for improvements in currently available retrovectors which may have the capability of transportation by natural transport systems in the human body will open effective ways for targeting human genes to specific cells in tissues in situ. Genetic engineering of virus vectors is a subject of prime importance to the developing gene therapy protocols in humans.Abbreviations HSV Herpes simplex virus     

Cytosine deaminase versus thymidine kinase: a comparison of the antitumor activity

The efficacy of single and combination suicide gene therapy was evaluated using a Herpes simplex virus thymidine kinase/ganciclovir system and Escherichia coli cytosine deaminase/5-fluorocytosine system on the rat prostate tumor cell line R3327 AT-1. The wild-type R3327 AT-1 cell line was transfected with a bifunctional fusion gene CDglyTK, which had the advantage that the resulting R3327 AT-1/CDglyTK cell line has the same amount of cytosine deaminase and thymidine kinase molecules. The percentage of viable R3327 AT-1/CDglyTK cells after 96 h incubation with 0.1 micro g/ml ganciclovir or 10 micro g/ml 5-fluorocytosine were 85% and 52% of controls, respectively. The cell viability when both suicide genes systems were activated was 43%. For in vivo analysis, Copenhagen rats were injected subcutaneously with R3327 AT-1 or R3327 AT-1/CDglyTK cells and treated with 30 mg/kg ganciclovir, 500 mg/kg 5-fluorocytosine, or both prodrugs together. A survival of 83% with the thymidine kinase/ganciclovir and 57% with the CD/5-FC could be observed. Only co-administration of thymidine kinase- and cytosine deaminase-specific prodrugs resulted in a 100% recurrence-free survival of the Copenhagen rats with a Dunning R3327 AT-1/CDglyTK prostate tumor and showed an additive cytotoxic effect. Calculation of the degree of activation and the potential of activation can be used to predict the success of a suicide gene therapy. In our case, the cytosine deaminase/5-fluorocytosine system had a low degree of activation (value 40), which is also found in the low response to 5- fluorocytosine in vivo (57% tumor free).     

Salvage of pyrimidine nucleosides by Trichomonas vaginalis

Trichomonas vaginalis is incapable of de novo pyrimidine biosynthesis because it cannot incorporate bicarbonate, aspartate or orotate into its pyrimidine nucleotides or nucleic acids. The organism can salvage exogenous cytidine greater than uridine greater than uracil and thymidine, and incorporate them into the nucleotide pool. A portion of cytidine is converted to CMP, CDP and CTP by cytidine phosphotransferase and nucleotide kinases. Some cytidine and most of uracil are, however, converted first to uridine by cytidine deaminase and uridine phosphorylase respectively; uridine is then incorporated into UMP, UDP and UTP by uridine phosphotransferase and nucleotide kinases. The two phosphotransferases, found mainly in the non-sedimentable fraction of T. vaginalis, provide the main avenue of pyrimidine salvage. No significant levels of pyrimidine phosphoribosyl transferase or nucleoside kinases can be detected in the extract. T. vaginalis has no appreciable dihydrofolate reductase or thymidylate synthetase; it grows normally in millimolar concentrations of methotrexate, pyrimethamine, or trimethoprim, and cannot incorporate labels from exogenous uracil or uridine into DNA. It has an enzyme thymidine phosphotransferase in the sedimentable fraction which converts thymidine to TMP. Thymidine salvage in T. vaginalis is thus totally isolated from the rest of the pyrimidine salvage.     

Prognostic factors in multiple myeloma: role of β2-microglobulin and thymidine kinase

Summary Serum ₂- microglobulin, serum thymidine kinase, and commonly used prognostic parameters were investigated for their prognostic value in a well-defined group of patients with multiple myeloma (n = 207). Multivariate analysis showed hemoglobin to be the parameter of strongest prognostic value. Only albumin, serum ₂-microglobulin and serum thymidine kinase added further prognostic information. When tested for efficiency in recognizing patients with poor (average survival time < 1 year) and good (average survival time > 5 years) prognosis, serum ₂-microglobulin was best (80%), followed by total urinary protein (78%), hemoglobin (76%), and albumin (75%).